Methods and compositions for the specific inhibition of alpha-1 antitrypsin by double-stranded RNA

ABSTRACT

This invention relates to compounds, compositions, and methods useful for reducing α-1 antitrypsin target RNA and protein levels via use of dsRNAs, e.g., Dicer substrate siRNA (DsiRNA) agents.

RELATED APPLICATIONS

The present application is a continuation of U.S. application Ser. No. 16/918,205, filed Jul. 1, 2020, which is a divisional of U.S. application Ser. No. 16/457,394, filed Jun. 28, 2019, which issued as U.S. Pat. No. 10,844,381 on Nov. 24, 2020, which is a divisional of U.S. patent application Ser. No. 15/247,201, filed on Aug. 25, 2016, which issued as U.S. Pat. No. 10,370,655 on Aug. 6, 2019, which is a divisional of U.S. patent application Ser. No. 14/323,299, filed on Jul. 3, 2014, which issued as U.S. Pat. No. 9,458,457 on Oct. 4, 2016, which claims priority to, and the benefit under 35 U.S.C. § 119(e) of the following applications: U.S. provisional patent application No. 61/842,551, entitled “Methods and Compositions for the Specific Inhibition of Alpha-1 Antitrypsin by Double-Stranded RNA,” filed Jul. 3, 2013; and U.S. provisional patent application No. 61/891,548, entitled “Methods and Compositions for the Specific Inhibition of Alpha-1 Antitrypsin by Double-Stranded RNA,” filed Oct. 16, 2013. The entire contents of the aforementioned patent applications are incorporated herein by this reference.

SEQUENCE SUBMISSION

The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is entitled Sequence Listing.txt, was created on Jul. 28, 2021, and is 689 kb in size. The information in the electronic format of the Sequence Listing is part of the present application and is incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to compounds, compositions, and methods for the study, diagnosis, and treatment of traits, diseases and conditions that respond to the modulation of α-1 antitrypsin gene expression and/or activity.

Sequence Submission

The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is entitled “301937_94007_seq_listing_1JUL2014”, was created on Jul. 1, 2014, and is 692 kb in size. The information in the electronic format of the Sequence Listing is part of the present application and is incorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION

Alpha 1-antitrypsin (AAT or Serpina1) is a protease inhibitor belonging to the serpin superfamily. It is generally known as serum trypsin inhibitor. Alpha 1-antitrypsin is also referred to as alpha-1 proteinase inhibitor (A1PI) because it inhibits a wide variety of proteases (Gettins PG. Chem Rev 102: 4751-804). It protects tissues from enzymes of inflammatory cells, especially neutrophil elastase, and has a reference range in blood of 1.5-3.5 gram/liter, but multi-fold elevated levels can occur upon acute inflammation (Kushner, Mackiewicz. Acute-phase glycoproteins: molecular biology, biochemistry and clinical applications (CRC Press). pp. 3-19). In the absence of AAT, neutrophil elastase is free to break down elastin, which contributes to the elasticity of the lungs, resulting in respiratory complications such as emphysema, or COPD (chronic obstructive pulmonary disease) in adults and cirrhosis in adults or children. Individuals with mutations in one or both copies of the AAT gene can suffer from alpha-1 anti-trypsin deficiency, which presents as a risk of developing pulmonary emphysema or chronic liver disease due to greater than normal elastase activity in the lungs and liver.

In affected individuals, the deficiency in alpha-1 antitrypsin is a deficiency of wildtype, functional alpha-1 antitrypsin. However, in some cases that are relevant to the current invention, the individual is producing significant quantities of alpha-1 antitrypsin, but a proportion of the alpha-1 antitrypsin protein being produced is misfolded or contains mutations that compromise the functioning of the protein. In some cases, the individual is producing misfolded proteins which cannot be properly transported from the site of synthesis to the site of action within the body.

Liver disease resulting from alpha-1 antitrypsin deficiency can be caused by such misfolded proteins. Mutant forms of alpha-1 antitypsin (e.g., the common PiZ variant, which harbors a glutamate to lysine mutation at position 342 (position 366 in pre-processed form)) are produced in liver cells (hepatocytes in the liver commonly produce a large amount of circulating AAT), and in the misfolded configuration, such forms are not readily transported out of the cells. This leads to a buildup of misfolded protein in the liver cells (hepatocytes, where those with the largest burden of mutant Z protein can suffer a cascade of intracellular damage that ultimately results in apoptosis; this chronic cycle of hepatocellular apoptosis and regeneration can lead to fibrosis and organ injury) and can cause one or more diseases or disorders of the liver including, but not limited to, chronic liver disease, liver inflammation, cirrhosis, liver fibrosis, and/or hepatocellular carcinoma.

There are currently few options for treating patients with liver disease associated with alpha-1 antitrypsin deficiency, and such options include hepatitis vaccination, supportive care, and avoidance of injurious agents (e.g., alcohol and NSAIDs), none of which provide a targeted therapy. Replacement of alpha-1 antitrypsin has no impact on liver disease in these patients but liver transplantation can be effective.

Double-stranded RNA (dsRNA) agents possessing strand lengths of 25 to 35 nucleotides have been described as effective inhibitors of target gene expression in mammalian cells (Rossi et al., U.S. Patent Application Nos. 2005/0244858 and US 2005/0277610). dsRNA agents of such length are believed to be processed by the Dicer enzyme of the RNA interference (RNAi) pathway, leading such agents to be termed “Dicer substrate siRNA” (“DsiRNA”) agents. Additional modified structures of DsiRNA agents were previously described (Rossi et al., U.S. Patent Application No. 2007/0265220). Effective extended forms of Dicer substrates have also recently been described (Brown, U.S. Pat. No. 8,349,809 and US 2010/0173974).

Provided herein are improved nucleic acid agents that target α-1 antitrypsin. In particular, those targeting α-1 antitrypsin have been specifically exemplified.

BRIEF SUMMARY OF THE INVENTION

The present invention is directed to nucleic acid compositions that reduce expression of α-1 antitrypsin. Such compositions contain nucleic acids such as double stranded RNA (“dsRNA”), and methods for preparing them. The nucleic acids of the invention are capable of reducing the expression of a target α-1 antitrypsin gene in a cell, either in vitro or in a mammalian subject.

In one aspect, the invention provides a nucleic acid having an oligonucleotide strand of 15-35 nucleotides in length that is sufficiently complementary to a target α-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 along at least 15 nucleotides of the oligonucleotide strand length to reduce α-1 antitrypsin target mRNA expression when the nucleic acid is introduced into a mammalian cell.

In another aspect, the invention provides a nucleic acid having an oligonucleotide strand of 19-35 nucleotides in length that is sufficiently complementary to a target α-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 along at least 19 nucleotides of the oligonucleotide strand length to reduce α-1 antitrypsin target mRNA expression when the nucleic acid is introduced into a mammalian cell.

In a further aspect, the invention provides a double stranded nucleic acid (dsNA) having first and second nucleic acid strands that include RNA, where the first strand is 15-35 nucleotides in length and the second strand is 19-35 nucleotides in length and is sufficiently complementary to a target α-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 along at least 15 nucleotides of the second oligonucleotide strand length to reduce α-1 antitrypsin target mRNA expression when the dsNA is introduced into a mammalian cell.

In an additional aspect, the invention provides a dsNA having first and second nucleic acid strands, where the first strand is 15-35 nucleotides in length and the second strand of the dsNA is 19-35 nucleotides in length and is sufficiently complementary to a target α-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 along at least 19 nucleotides of the second oligonucleotide strand length to reduce α-1 antitrypsin target mRNA expression when the dsNA is introduced into a mammalian cell.

In another aspect, the invention provides a dsNA having first and second nucleic acid strands, where the first strand is 15-35 nucleotides in length and the second strand of the dsNA is 19-35 nucleotides in length and is sufficiently complementary to a target α-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 along at least 19 nucleotides of the second oligonucleotide strand length to reduce α-1 antitrypsin target mRNA expression, and where, starting from the 5′ end of the α-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 (referred to as position 1), mammalian Ago2 cleaves the mRNA at a site between positions 9 and 10 of the sequence when the dsNA is introduced into a mammalian cell.

In a further aspect, the invention provides a dsNA molecule that consists of (a) a sense region and an antisense region, where the sense region and the antisense region together form a duplex region consisting of 25-35 base pairs and the antisense region comprises a sequence that is the complement of a sequence of any one (or more) of SEQ ID NOs: 991-1188 and 1938-1968; and (b) from zero to two 3′ overhang regions, where each overhang region is six or fewer nucleotides in length, and where, starting from the 5′ end of the α-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 and 1938-1968 (position 1), mammalian Ago2 cleaves the mRNA at a site between positions 9 and 10 of the sequence when the dsNA is introduced into a mammalian cell.

In an additional aspect, the invention provides a dsNA having first and second nucleic acid strands and a duplex region of at least 25 base pairs, where the first strand is 25-34 nucleotides in length and the second strand of the dsNA is 26-35 nucleotides in length and includes 1-5 single-stranded nucleotides at its 3′ terminus, where the second oligonucleotide strand is sufficiently complementary to a target α-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 and 1938-1968 along at least 19 nucleotides of the second oligonucleotide strand length to reduce α-1 antitrypsin target gene expression when the dsNA is introduced into a mammalian cell.

In another aspect, the invention provides a dsNA having first and second nucleic acid strands and a duplex region of at least 25 base pairs, where the first strand is 25-34 nucleotides in length and the second strand of the dsNA is 26-35 nucleotides in length and comprises 1-5 single-stranded nucleotides at its 3′ terminus, where the 3′ terminus of the first oligonucleotide strand and the 5′ terminus of the second oligonucleotide strand form a blunt end, and the second oligonucleotide strand is sufficiently complementary to a target α-1 antitrypsin sequence of SEQ ID NOs: 991-1188 and 1938-1968 along at least 19 nucleotides of the second oligonucleotide strand length to reduce α-1 antitrypsin mRNA expression when the dsNA is introduced into a mammalian cell.

In an additional aspect, the invention provides a nucleic acid possessing an oligonucleotide strand of 15-35 nucleotides in length, where the oligonucleotide strand is hybridizable to a target α-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 or 1938-1968 along at least 15 nucleotides of the oligonucleotide strand length.

Another aspect of the invention provides a dsNA having first and second nucleic acid strands that include RNA, where the first strand is 15-35 nucleotides in length and the second strand of the dsNA is 19-35 nucleotides in length, where the second oligonucleotide strand is hybridizable to a target α-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 or 1938-1968 along at least 15 nucleotides of the second oligonucleotide strand length.

A further aspect of the invention provides an in vivo hybridization complex within a cell that includes an exogenous nucleic acid sequence and a target alpha-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 or 1938-1968.

An additional aspect of the invention provides an in vitro hybridization complex within a cell that includes an exogenous nucleic acid sequence and a target alpha-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 or 1938-1968.

In one embodiment, the dsNA has a duplex region that is 19-21 base pairs, 21-25 base pairs or at least 25 base pairs in length.

In another embodiment, the second oligonucleotide strand includes 1-5 single-stranded nucleotides at its 3′ terminus.

In an additional embodiment, the first strand is 25-35 nucleotides in length. Optionally, the second strand is 25-35 nucleotides in length.

In another embodiment, the second oligonucleotide strand is complementary to target α-1 antitrypsin cDNA sequence GenBank Accession No. NM_000295.4 along at most 27 nucleotides of the second oligonucleotide strand length.

In one embodiment, the dsNA or hybridization complex comprises a modified nucleotide. Optionally, the modified nucleotide residue is of the group consisting of 2′-O-methyl, 2′-methoxyethoxy, 2′-fluoro, 2′-allyl, 2′-O-[2-(methylamino)-2-oxoethyl], 4′-thio, 4′-CH2-O-2′-bridge, 4′-(CH2)2-O-2′-bridge, 2′-LNA, 2′-amino and 2′-O-(N-methlycarbamate).

In a further embodiment, starting from the first nucleotide (position 1) at the 3′ terminus of the first oligonucleotide strand, position 1, 2 or 3 is substituted with a modified nucleotide. Optionally, the modified nucleotide residue of the 3′ terminus of the first strand is a deoxyribonucleotide, an acyclonucleotide or a fluorescent molecule. In certain embodiments, position 1 of the 3′ terminus of the first oligonucleotide strand is a deoxyribonucleotide.

In one embodiment, the first strand is 25 nucleotides in length and the second strand is 27 nucleotides in length. Optionally, the 3′ terminus of the first strand and the 5′ terminus of the second strand form a blunt end.

In another embodiment, starting from the 5′ end of a α-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 and 1938-1968 (position 1), mammalian Ago2 cleaves the mRNA at a site between positions 9 and 10 of the sequence, thereby reducing α-1 antitrypsin target mRNA expression when the dsNA is introduced into a mammalian cell. Optionally, the second strand includes a sequence of SEQ ID NOs: 199-396 or 3493-3499. In certain embodiments, the first strand includes a sequence of SEQ ID NOs: 1-198.

In one embodiment, the dsNA includes a pair of first strand/second strand sequences of Table 2.

In a further embodiment, each of the first and the second strands has a length which is at least 26 nucleotides.

In one embodiment, nucleotides of the 1-5 single-stranded nucleotides of the 3′ terminus of the second strand include a modified nucleotide. Optionally, the modified nucleotide is a 2′-O-methyl ribonucleotide. In a related embodiment, all nucleotides of the 1-5 single-stranded nucleotides of the 3′ terminus of the second strand are modified nucleotides. In certain embodiments, the 1-5 single-stranded nucleotides of the 3′ terminus of the second strand are 1-4 nucleotides in length, are 1-3 nucleotides in length, or are 1-2 nucleotides in length. In a related embodiment, the 1-5 single-stranded nucleotides of the 3′ terminus of the second strand is two nucleotides in length and includes a 2′-O-methyl modified ribonucleotide.

In certain embodiments, the second oligonucleotide strand includes a modification pattern of AS-M1 to AS-M84, AS-M88 to AS-M96, AS-M210, AS-M1* to AS-M84*, AS-M88* to AS-M96* or AS-M210*.

Optionally, the first oligonucleotide strand includes a modification pattern of SM1 to SM119 or SM250 to SM252.

In a further embodiment, each of the first and the second strands has a length which is at least 26 and at most 30 nucleotides.

Optionally, the dsNA is cleaved endogenously in the cell by Dicer.

In certain embodiments, a dsNA of the invention includes at least one unlocked nucleobase analog (UNA). Optionally, the at least one UNA is located in a 3′-overhang region, a 5′-overhang region, or both such regions of the dsNA, optionally on the guide strand of the dsNA.

In some embodiments, a dsNA of the invention is attached to a dynamic polyconjugate (DPC). In additional embodiments, a dsNA of the invention is administered with a DPC, where optionally the dsNA and DPC are not attached.

In some embodiments, a dsNA of the invention is attached to a GalNAc moiety (optionally, a tri-antennary GalNAc moiety) and/or to cholesterol or a cholesterol targeting ligand.

In certain embodiments, the amount of the nucleic acid sufficient to reduce expression of the target gene is 1 nanomolar or less, 200 picomolar or less, 100 picomolar or less, 50 picomolar or less, 20 picomolar or less, 10 picomolar or less, 5 picomolar or less, 2, picomolar or less or 1 picomolar or less in the environment of the cell.

In one embodiment, the dsNA possesses greater potency than a 21mer siRNA directed to the identical at least 19 nucleotides of the target α-1 antitrypsin mRNA in reducing target α-1 antitrypsin mRNA expression when assayed in vitro in a mammalian cell at an effective concentration in the environment of a cell of 1 nanomolar or less. In certain embodiments, knockdown efficacy and/or potency is measured at a concentration of 1 nanomolar, 200 picomolar, 100 picomolar, 50 picomolar, 20 picomolar, 10 picomolar, 5 picomolar, 2, picomolar or 1 picomolar in the environment of a cell.

In another embodiment, the dsNA is sufficiently complementary to the target α-1 antitrypsin mRNA sequence to reduce α-1 antitrypsin target mRNA expression by an amount (expressed by %) that is at least 10%, at least 50%, at least 80-90%, at least 95%, at least 98%, or at least 99% when the dsNA is introduced into a mammalian cell.

In certain embodiments, the first and second strands are joined by a chemical linker. Optionally, the 3′ terminus of the first strand and the 5′ terminus of the second strand are joined by a chemical linker.

In one embodiment, a nucleotide of the second or first strand is substituted with a modified nucleotide that directs the orientation of Dicer cleavage.

Optionally, the dsNA includes a deoxyribonucleotide, a dideoxyribonucleotide, an acyclonucleotide, a 3′-deoxyadenosine (cordycepin), a 3′-azido-3′-deoxythymidine (AZT), a 2′,3′-dideoxyinosine (ddI), a 2′,3′-dideoxy-3′-thiacytidine (3TC), a 2′,3′-didehydro-2′,3′-dideoxythymidine (d4T), a monophosphate nucleotide of 3′-azido-3′-deoxythymidine (AZT), a 2′,3′-dideoxy-3′-thiacytidine (3TC) and a monophosphate nucleotide of 2′,3′-didehydro-2′,3′-dideoxythymidine (d4T), a 4-thiouracil, a 5-bromouracil, a 5-iodouracil, a 5-(3-aminoallyl)-uracil, a 2′-O-alkyl ribonucleotide, a 2′-O-methyl ribonucleotide, a 2′-amino ribonucleotide, a 2′-fluoro ribonucleotide, or a locked nucleic acid.

In certain embodiments, the dsNA includes a phosphate backbone modification that is a phosphonate, a phosphorothioate or a phosphotriester.

In one embodiment, the dsNA includes a morpholino nucleic acid or a peptide nucleic acid (PNA).

In one aspect, the invention provides a method for reducing expression of a target α-1 antitrypsin gene in a mammalian cell involving contacting a mammalian cell in vitro with a dsNA of the invention in an amount sufficient to reduce expression of a target α-1 antitrypsin mRNA in the cell.

In one embodiment, target α-1 antitrypsin mRNA expression is reduced by at least 10%, at least 50% or at least 80-90%. Optionally, α-1 antitrypsin mRNA levels are reduced by at least 90% at least 8 days after the cell is contacted with the dsNA. In certain embodiments, α-1 antitrypsin mRNA levels are reduced by at least 70% at least 10 days after the cell is contacted with the dsNA.

In one aspect, the invention provides a method for reducing expression of a target α-1 antitrypsin mRNA in a mammal involving administering a nucleic acid of the invention to a mammal in an amount sufficient to reduce expression of a target α-1 antitrypsin mRNA in the mammal.

In certain embodiments, the nucleic acid is formulated in a lipid nanoparticle (LNP). In one embodiment, the nucleic acid is administered at a dosage that is 1 microgram to 5 milligrams per kilogram of the mammal per day, 100 micrograms to 0.5 milligrams per kilogram, 0.001 to 0.25 milligrams per kilogram, 0.01 to 20 micrograms per kilogram, 0.01 to 10 micrograms per kilogram, 0.10 to 5 micrograms per kilogram, or 0.1 to 2.5 micrograms per kilogram.

In certain embodiments, the nucleic acid possesses greater potency than 21mer siRNAs directed to the identical at least 19 nucleotides of the target α-1 antitrypsin mRNA in reducing target α-1 antitrypsin mRNA expression when assayed in vitro in a mammalian cell at an effective concentration in the environment of a cell of 1 nanomolar or less. In certain embodiments, knockdown efficacy and/or potency is measured at a concentration of 1 nanomolar, 200 picomolar, 100 picomolar, 50 picomolar, 20 picomolar, 10 picomolar, 5 picomolar, 2, picomolar or 1 picomolar in the environment of a cell.

In another embodiment, α-1 antitrypsin mRNA levels are reduced in a tissue of the mammal by at least 70% at least 3 days after the dsNA is administered to the mammal. Optionally, the tissue is liver tissue.

In certain embodiments, administering includes intravenous injection, intramuscular injection, intraperitoneal injection, infusion, subcutaneous injection, transdermal, aerosol, rectal, vaginal, topical, oral or inhaled delivery.

In another aspect, the invention provides a method for treating or preventing a liver disease or disorder in a subject that involves administering a dsNA of the invention to the subject in an amount sufficient to treat or prevent the liver disease or disorder in the subject.

In one embodiment, the liver disease or disorder is a chronic liver disease, liver inflammation, cirrhosis, liver fibrosis or hepatocellular carcinoma. Optionally, the subject is human.

In a further aspect, the invention provides a formulation containing a nucleic acid of the invention present in an amount effective to reduce target α-1 antitrypsin mRNA levels when the nucleic acid is introduced into a mammalian cell in vitro by at least 10%, at least 50% or at least 80-90%.

In one embodiment, the effective amount is 1 nanomolar or less, 200 picomolar or less, 100 picomolar or less, 50 picomolar or less, 20 picomolar or less, 10 picomolar or less, 5 picomolar or less, 2, picomolar or less or 1 picomolar or less in the environment of the cell. In certain embodiments, knockdown efficacy is measured at a concentration of 1 nanomolar, 200 picomolar, 100 picomolar, 50 picomolar, 20 picomolar, 10 picomolar, 5 picomolar, 2, picomolar or 1 picomolar in the environment of a cell.

In another aspect, the invention provides a formulation including the dsNA of the invention present in an amount effective to reduce target α-1 antitrypsin mRNA levels when the dsNA is introduced into a cell of a mammalian subject by at least 10%, at least 50% or at least 80-90%. Optionally, the effective amount is a dosage of 1 microgram to 5 milligrams per kilogram of the subject per day, 100 micrograms to 0.5 milligrams per kilogram, 0.001 to 0.25 milligrams per kilogram, 0.01 to 20 micrograms per kilogram, 0.01 to 10 micrograms per kilogram, 0.10 to 5 micrograms per kilogram, or 0.1 to 2.5 micrograms per kilogram.

In an additional aspect, the invention provides a mammalian cell containing a nucleic acid of the invention.

In a further aspect, the invention provides a pharmaceutical composition containing a nucleic acid of the invention and a pharmaceutically acceptable carrier.

In another aspect, the invention provides a kit that includes a nucleic acid of the invention and instructions for its use.

In an additional aspect, the invention provides a composition possessing α-1 antitrypsin inhibitory activity that consists essentially of a nucleic acid of the invention.

Another aspect of the invention provides a method of hybridizing an exogenous nucleic acid to mRNA in a cell that involves introducing into the cell an exogenous nucleic acid sequence and hybridizing the exogenous nucleic acid to a target alpha-1 antitrypsin mRNA sequence that is a sequence of SEQ ID NOs: 991-1188 or 1938-1968.

A further aspect of the invention provides a method of treating an individual with a liver or lung disease or disorder that involves introducing into cells of the individual an exogenous nucleic acid sequence and hybridizing the exogenous nucleic acid to a target alpha-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 or 1938-1968.

An additional aspect of the invention provides a method of forming an in vivo hybridization complex within a cell that involves introducing into the cell an exogenous nucleic acid sequence and hybridizing the exogenous nucleic acid to a target alpha-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 or 1938-1968.

A further aspect of the invention provides a method of inhibiting translation of a target mRNA into a protein within a cell that involves introducing into the cell an exogenous nucleic acid sequence and hybridizing the exogenous nucleic acid to a target alpha-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 or 1938-1968, complexing the exogenous nucleic acid with RISC, and cleaving the mRNA.

In one embodiment, the exogenous nucleic acid is complexed with RISC. In a related embodiment, the RISC cleaves the mRNA.

Another aspect of the invention provides a “single strand-extended” (here, guide strand-extended, at the 5′ end) dsNA. Specifically, this aspect of the invention provides a dsNA having first and second nucleic acid strands that include RNA, where the first strand is 15-35 nucleotides in length and the second strand is at least 35 nucleotides in length, optionally includes a second strand sequence having at least 25 nucleotides of a sequence of SEQ ID NOs: 3493-3499 and is sufficiently complementary to a target α-1 antitrypsin mRNA sequence of SEQ ID NOs: 991-1188 along at least 15 nucleotides of the second oligonucleotide strand length to reduce α-1 antitrypsin target mRNA expression when the dsNA is introduced into a mammalian cell.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the structures of exemplary DsiRNA agents of the invention targeting a site in the α-1 antitrypsin RNA referred to herein as the “α-1 antitrypsin-506” target site. UPPER case=unmodified RNA, lower case=DNA, Bold=mismatch base pair nucleotides; arrowheads indicate projected Dicer enzyme cleavage sites; dashed line indicates sense strand (top strand) sequences corresponding to the projected Argonaute 2 (Ago2) cleavage site within the targeted α-1 antitrypsin sequence.

FIGS. 2A to 2D present primary screen data showing DsiRNA-mediated knockdown of human α-1 antitrypsin (FIGS. 2A and 2B) and mouse α-1 antitrypsin (FIGS. 2C and 2D) in human (Huh7) and mouse (AML12) cells, respectively. For each DsiRNA tested, two independent qPCR amplicons were assayed (in human cells, amplicons “462-563” and “1811-1910” were assayed, while in mouse cells, amplicons “331-462” and “1532-1662” were assayed).

FIGS. 3A to 3H show histograms of human and mouse α-1 antitrypsin inhibitory efficacies observed for indicated DsiRNAs. “P1” indicates phase 1 (primary screen), while “P2” indicates phase 2. In phase 1, DsiRNAs were tested at 1 nM in the environment of Huh7 cells (human cell assays; FIGS. 3A to 3D) or mouse cells (AML12 cell assays; FIGS. 3E to 3H). In phase 2, DsiRNAs were tested at 1 nM, 0.1 nM and 0.03 nM in the environment of Huh7 cells. Individual bars represent average human (FIGS. 3A to 3D) or mouse (FIGS. 3E to 3H) α-1 antitrypsin levels observed in triplicate, with standard errors shown. Human α-1 antitrypsin levels were normalized to HPRT and SFRS9 levels, while mouse α-1 antitrypsin levels were normalized to HPRT and Rpl23 levels.

FIGS. 4A to 4F present data showing levels of α-1 antitrypsin knockdown observed for 24 α-1 antitrypsin-targeting duplex sequences possessing a range of guide strand 2′-O-methyl modifications and a single passenger strand 2′-O-methyl modification pattern, as depicted in FIGS. 4A and 4B. Bar graphs of FIGS. 4C to 4F show efficacy data for the 24 independent α-1 antitrypsin-targeting DsiRNAs across different, indicated guide (antisense) strand 2′-O-methyl modification patterns in human Huh7 cells at 0.03 nM, 0.1 nM and at 1 nM.

FIGS. 5A to 5H present data obtained in an expanded modified duplex screen. FIGS. 5A to 5D depict 2′-O-methyl modification patterns of both passenger and guide strands of tested duplexes (with FIG. 5D showing modification patterns of individual strands), while FIGS. 5E to 5H show histograms of human α-1 antitrypsin inhibitory efficacies observed for indicated DsiRNAs in human cells. “P4” indicates phase 4 (expanded modified duplex screen). In the expanded modification screen, DsiRNAs were tested at 0.03 nM, 0.1 nM and at 1 nM in the environment of human Huh7 cells. Individual bars represent average human α-1 antitrypsin levels observed in triplicate, with standard errors shown. Human α-1 antitrypsin levels were normalized to HPRT and SFRS9 levels.

FIGS. 6A and 6B demonstrate the robust α-1 antitrypsin knockdown efficacy of guide strand “extended” forms of α-1 antitrypsin-targeting DsiRNAs of the invention. FIG. 6A shows the specific sequences of each of the eight extended DsiRNAs tested (corresponding SEQ ID NOs are indicated at right for sense and antisense sequences, respectively). FIG. 6B shows IC₅₀ curves obtained for each of the eight guide strand “extended” forms of α-1 antitrypsin-targeting DsiRNAs that were tested (“C121” represents a control DsiRNA that does not target α-1 antitrypsin).

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to compositions that contain nucleic acids, for example double stranded RNA (“dsRNA”), and methods for preparing them, that are capable of reducing the level and/or expression of the α-1 antitrypsin gene in vivo or in vitro. One of the strands of the dsRNA contains a region of nucleotide sequence that has a length that ranges from 19 to 35 nucleotides that can direct the destruction and/or translational inhibition of the targeted α-1 antitrypsin transcript.

Definitions

Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which this invention belongs. The following references provide one of skill with a general definition of many of the terms used in this invention: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used herein, the following terms have the meanings ascribed to them below, unless specified otherwise.

The present invention features one or more DsiRNA molecules that can modulate (e.g., inhibit) α-1 antitrypsin expression. The DsiRNAs of the invention optionally can be used in combination with modulators of other genes and/or gene products associated with the maintenance or development of diseases or disorders associated with α-1 antitrypsin misregulation (e.g., tumor formation and/or growth, etc.). The DsiRNA agents of the invention modulate α-1 antitrypsin RNAs such as those corresponding to the cDNA sequences referred to by GenBank Accession Nos. NM_000295.4 (human α-1 antitrypsin) and NM_009246.3 (mouse Serpina1d), which are referred to herein generally as “α-1 antitrypsin.”

The below description of the various aspects and embodiments of the invention is provided with reference to exemplary α-1 antitrypsin RNAs, generally referred to herein as α-1 antitrypsin. However, such reference is meant to be exemplary only and the various aspects and embodiments of the invention are also directed to alternate α-1 antitrypsin RNAs, such as mutant α-1 antitrypsin RNAs or additional α-1 antitrypsin splice variants. Certain aspects and embodiments are also directed to other genes involved in α-1 antitrypsin pathways, including genes whose misregulation acts in association with that of α-1 antitrypsin (or is affected or affects α-1 antitrypsin regulation) to produce phenotypic effects that may be targeted for treatment (e.g., tumor formation and/or growth, etc.). BAK1, Noxa, BCL2L11, Bc1-2-associated death promoter, PCNA, DAD1, TNKS and BH3 interacting domain death agonist are examples of genes that interact with α-1 antitrypsin. Such additional genes, including those of pathways that act in coordination with α-1 antitrypsin, can be targeted using dsRNA and the methods described herein for use of α-1 antitrypsin-targeting dsRNAs. Thus, the inhibition and the effects of such inhibition of the other genes can be performed as described herein.

The term “α-1 antitrypsin” refers to nucleic acid sequences encoding a α-1 antitrypsin protein, peptide, or polypeptide (e.g., α-1 antitrypsin transcripts, such as the sequences of α-1 antitrypsin Genbank Accession Nos. NM_000295.4 and NM_009246.3). In certain embodiments, the term “α-1 antitrypsin” is also meant to include other α-1 antitrypsin encoding sequence, such as other α-1 antitrypsin isoforms, mutant α-1 antitrypsin genes, splice variants of α-1 antitrypsin genes, and α-1 antitrypsin gene polymorphisms. The term “α-1 antitrypsin” is also used to refer to the polypeptide gene product of an α-1 antitrypsin gene/transcript, e.g., an α-1 antitrypsin protein, peptide, or polypeptide, such as those encoded by α-1 antitrypsin Genbank Accession Nos. NP_000286.3 and NP_033272.1.

As used herein, a “α-1 antitrypsin-associated disease or disorder” refers to a disease or disorder known in the art to be associated with altered α-1 antitrypsin expression, level and/or activity. Notably, a “α-1 antitrypsin-associated disease or disorder” includes diseases or disorders of the liver including, but not limited to, chronic liver disease, liver inflammation, cirrhosis, liver fibrosis, and/or hepatocellular carcinoma.

An anti-α-1 antitrypsin dsRNA of the invention is deemed to possess “α-1 antitrypsin inhibitory activity” if a statistically significant reduction in α-1 antitrypsin RNA (or when the α-1 antitrypsin protein is assessed, α-1 antitrypsin protein levels) is seen when an anti-α-1 antitrypsin dsRNA of the invention is administered to a system (e.g., cell-free in vitro system), cell, tissue or organism, as compared to a selected control. The distribution of experimental values and the number of replicate assays performed will tend to dictate the parameters of what levels of reduction in α-1 antitrypsin RNA (either as a % or in absolute terms) is deemed statistically significant (as assessed by standard methods of determining statistical significance known in the art). However, in certain embodiments, “α-1 antitrypsin inhibitory activity” is defined based upon a % or absolute level of reduction in the level of α-1 antitrypsin in a system, cell, tissue or organism. For example, in certain embodiments, a dsRNA of the invention is deemed to possess α-1 antitrypsin inhibitory activity if at least a 5% reduction or at least a 10% reduction in α-1 antitrypsin RNA is observed in the presence of a dsRNA of the invention relative to α-1 antitrypsin levels seen for a suitable control. (For example, in vivo α-1 antitrypsin levels in a tissue and/or subject can, in certain embodiments, be deemed to be inhibited by a dsRNA agent of the invention if, e.g., a 5% or 10% reduction in α-1 antitrypsin levels is observed relative to a control.) In certain other embodiments, a dsRNA of the invention is deemed to possess α-1 antitrypsin inhibitory activity if α-1 antitrypsin RNA levels are observed to be reduced by at least 15% relative to a selected control, by at least 20% relative to a selected control, by at least 25% relative to a selected control, by at least 30% relative to a selected control, by at least 35% relative to a selected control, by at least 40% relative to a selected control, by at least 45% relative to a selected control, by at least 50% relative to a selected control, by at least 55% relative to a selected control, by at least 60% relative to a selected control, by at least 65% relative to a selected control, by at least 70% relative to a selected control, by at least 75% relative to a selected control, by at least 80% relative to a selected control, by at least 85% relative to a selected control, by at least 90% relative to a selected control, by at least 95% relative to a selected control, by at least 96% relative to a selected control, by at least 97% relative to a selected control, by at least 98% relative to a selected control or by at least 99% relative to a selected control. In some embodiments, complete inhibition of α-1 antitrypsin is required for a dsRNA to be deemed to possess α-1 antitrypsin inhibitory activity. In certain models (e.g., cell culture), a dsRNA is deemed to possess α-1 antitrypsin inhibitory activity if at least a 50% reduction in α-1 antitrypsin levels is observed relative to a suitable control. In certain other embodiments, a dsRNA is deemed to possess α-1 antitrypsin inhibitory activity if at least an 80% reduction in α-1 antitrypsin levels is observed relative to a suitable control.

By way of specific example, in Example 2 below, a series of DsiRNAs targeting α-1 antitrypsin were tested for the ability to reduce α-1 antitrypsin mRNA levels in human Huh7 or mouse AML12 cells in vitro, at 1 nM concentrations in the environment of such cells and in the presence of a transfection agent (Lipofectamine™ RNAiMAX, Invitrogen). Within Example 2 below, α-1 antitrypsin inhibitory activity was ascribed to those DsiRNAs that were observed to effect at least a 70% reduction of α-1 antitrypsin mRNA levels under the assayed conditions. It is contemplated that α-1 antitrypsin inhibitory activity could also be attributed to a dsRNA under either more or less stringent conditions than those employed for Example 2 below, even when the same or a similar assay and conditions are employed. For example, in certain embodiments, a tested dsRNA of the invention is deemed to possess α-1 antitrypsin inhibitory activity if at least a 10% reduction, at least a 20% reduction, at least a 30% reduction, at least a 40% reduction, at least a 50% reduction, at least a 60% reduction, at least a 75% reduction, at least an 80% reduction, at least an 85% reduction, at least a 90% reduction, or at least a 95% reduction in α-1 antitrypsin mRNA levels is observed in a mammalian cell line in vitro at 1 nM dsRNA concentration or lower in the environment of a cell, relative to a suitable control.

Use of other endpoints for determination of whether a double stranded RNA of the invention possesses α-1 antitrypsin inhibitory activity is also contemplated. Specifically, in one embodiment, in addition to or as an alternative to assessing α-1 antitrypsin mRNA levels, the ability of a tested dsRNA to reduce α-1 antitrypsin protein levels (e.g., at 48 hours after contacting a mammalian cell in vitro or in vivo) is assessed, and a tested dsRNA is deemed to possess α-1 antitrypsin inhibitory activity if at least a 10% reduction, at least a 20% reduction, at least a 30% reduction, at least a 40% reduction, at least a 50% reduction, at least a 60% reduction, at least a 70% reduction, at least a 75% reduction, at least an 80% reduction, at least an 85% reduction, at least a 90% reduction, or at least a 95% reduction in α-1 antitrypsin protein levels is observed in a mammalian cell contacted with the assayed double stranded RNA in vitro or in vivo, relative to a suitable control. Additional endpoints contemplated include, e.g., assessment of a phenotype associated with reduction of α-1 antitrypsin levels—e.g., reduction of chronic liver disease, liver inflammation, cirrhosis, liver fibrosis, and/or hepatocellular carcinoma, as assessed directly or via assessment of appropriate markers and/or indicators of such liver disease or disorder.

α-1 antitrypsin inhibitory activity can also be evaluated over time (duration) and over concentration ranges (potency), with assessment of what constitutes a dsRNA possessing α-1 antitrypsin inhibitory activity adjusted in accordance with concentrations administered and duration of time following administration. Thus, in certain embodiments, a dsRNA of the invention is deemed to possess α-1 antitrypsin inhibitory activity if at least a 50% reduction in α-1 antitrypsin activity is observed/persists at a duration of time of 2 hours, 5 hours, 10 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days or more after administration of the dsRNA to a cell or organism. In additional embodiments, a dsRNA of the invention is deemed to be a potent α-1 antitrypsin inhibitory agent if α-1 antitrypsin inihibitory activity (e.g., in certain embodiments, at least 50% inhibition of α-1 antitrypsin) is observed at a concentration of 1 nM or less, 500 pM or less, 200 pM or less, 100 pM or less, 50 pM or less, 20 pM or less, 10 pM or less, 5 pM or less, 2 pM or less or even 1 pM or less in the environment of a cell, for example, within an in vitro assay for α-1 antitrypsin inihibitory activity as described herein. In certain embodiments, a potent α-1 antitrypsin inhibitory dsRNA of the invention is defined as one that is capable of α-1 antitrypsin inihibitory activity (e.g., in certain embodiments, at least 20% reduction of α-1 antitrypsin levels) at a formulated concentration of 10 mg/kg or less when administered to a subject in an effective delivery vehicle (e.g., an effective lipid nanoparticle formulation). Preferably, a potent α-1 antitrypsin inhibitory dsRNA of the invention is defined as one that is capable of α-1 antitrypsin inihibitory activity (e.g., in certain embodiments, at least 50% reduction of α-1 antitrypsin levels) at a formulated concentration of 5 mg/kg or less when administered to a subject in an effective delivery vehicle. More preferably, a potent α-1 antitrypsin inhibitory dsRNA of the invention is defined as one that is capable of α-1 antitrypsin inihibitory activity (e.g., in certain embodiments, at least 50% reduction of α-1 antitrypsin levels) at a formulated concentration of 5 mg/kg or less when administered to a subject in an effective delivery vehicle. Optionally, a potent α-1 antitrypsin inhibitory dsRNA of the invention is defined as one that is capable of α-1 antitrypsin inhibitory activity (e.g., in certain embodiments, at least 50% reduction of α-1 antitrypsin levels) at a formulated concentration of 2 mg/kg or less, or even 1 mg/kg or less, when administered to a subject in an effective delivery vehicle. Exemplary discrete formulated concentrations of α-1 antitrypsin-targeting RNAi agents of the invention include about 5 mg/kg, about 2 mg/kg, about 1 mg/kg, about 500 mg/kg, about 250 mg/kg, about 100 mg/kg, about 50 mg/kg, about 25 mg/kg, about 10 μg/kg, about 5 μg/kg, about 2.5 μg/kg, about 1 μg/kg, about 500 ng/kg, about 250 ng/kg, about 100 ng/kg, about 50 ng/kg, about 25 ng/kg, about 10 ng/kg, about 5 ng/kg, about 2.5 ng/kg, about 1 ng/kg and about 500 pg/kg.

About: As used herein, the term “about” means+/−10% of the recited value. Use of “about” is contemplated in reference to all ranges and values recited herein.

In certain embodiments, the phrase “consists essentially of” is used in reference to the anti-α-1 antitrypsin dsRNAs of the invention. In some such embodiments, “consists essentially of” refers to a composition that comprises a dsRNA of the invention which possesses at least a certain level of α-1 antitrypsin inhibitory activity (e.g., at least 50% α-1 antitrypsin inhibitory activity) and that also comprises one or more additional components and/or modifications that do not significantly impact the α-1 antitrypsin inhibitory activity of the dsRNA. For example, in certain embodiments, a composition “consists essentially of” a dsRNA of the invention where modifications of the dsRNA of the invention and/or dsRNA-associated components of the composition do not alter the α-1 antitrypsin inhibitory activity (optionally including potency or duration of α-1 antitrypsin inhibitory activity) by greater than 3%, greater than 5%, greater than 10%, greater than 15%, greater than 20%, greater than 25%, greater than 30%, greater than 35%, greater than 40%, greater than 45%, or greater than 50% relative to the dsRNA of the invention in isolation. In certain embodiments, a composition is deemed to consist essentially of a dsRNA of the invention even if more dramatic reduction of α-1 antitrypsin inhibitory activity (e.g., 80% reduction, 90% reduction, etc. in efficacy, duration and/or potency) occurs in the presence of additional components or modifications, yet where α-1 antitrypsin inhibitory activity is not significantly elevated (e.g., observed levels of α-1 antitrypsin inhibitory activity are within 10% those observed for the dsRNA of the invention) in the presence of additional components and/or modifications.

As used herein, the term “nucleic acid” refers to deoxyribonucleotides, ribonucleotides, or modified nucleotides, and polymers thereof in single- or double-stranded form. The term encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides. Examples of such analogs include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs) and unlocked nucleic acids (UNAs or unlocked nucleobase analogs; see, e.g., Jensen et al. Nucleic Acids Symposium Series 52: 133-4), and derivatives thereof.

As used herein, “nucleotide” is used as recognized in the art to include those with natural bases (standard), and modified bases well known in the art. Such bases are generally located at the 1′ position of a nucleotide sugar moiety. Nucleotides generally comprise a base, sugar and a phosphate group. The nucleotides can be unmodified or modified at the sugar, phosphate and/or base moiety, (also referred to interchangeably as nucleotide analogs, modified nucleotides, non-natural nucleotides, non-standard nucleotides and other; see, e.g., Usman and McSwiggen, supra; Eckstein, et al., International PCT Publication No. WO 92/07065; Usman et al, International PCT Publication No. WO 93/15187; Uhlman & Peyman, supra, all are hereby incorporated by reference herein). There are several examples of modified nucleic acid bases known in the art as summarized by Limbach, et al, Nucleic Acids Res. 22:2183, 1994. Some of the non-limiting examples of base modifications that can be introduced into nucleic acid molecules include, hypoxanthine, purine, pyridin-4-one, pyridin-2-one, phenyl, pseudouracil, 2,4,6-trimethoxy benzene, 3-methyl uracil, dihydrouridine, naphthyl, aminophenyl, 5-alkylcytidines (e.g., 5-methylcytidine), 5-alkyluridines (e.g., ribothymidine), 5-halouridine (e.g., 5-bromouridine) or 6-azapyrimidines or 6-alkylpyrimidines (e.g. 6-methyluridine), propyne, and others (Burgin, et al., Biochemistry 35:14090, 1996; Uhlman & Peyman, supra). By “modified bases” in this aspect is meant nucleotide bases other than adenine, guanine, cytosine and uracil at 1′ position or their equivalents.

As used herein, “modified nucleotide” refers to a nucleotide that has one or more modifications to the nucleoside, the nucleobase, pentose ring, or phosphate group. For example, modified nucleotides exclude ribonucleotides containing adenosine monophosphate, guanosine monophosphate, uridine monophosphate, and cytidine monophosphate and deoxyribonucleotides containing deoxyadenosine monophosphate, deoxyguanosine monophosphate, deoxythymidine monophosphate, and deoxycytidine monophosphate. Modifications include those naturally occurring that result from modification by enzymes that modify nucleotides, such as methyltransferases. Modified nucleotides also include synthetic or non-naturally occurring nucleotides. Synthetic or non-naturally occurring modifications in nucleotides include those with 2′ modifications, e.g., 2′-methoxyethoxy, 2′-fluoro, 2′-allyl, 2′-O-[2-(methylamino)-2-oxoethyl], 4′-thio, 4′-CH₂—O-2′-bridge, 4′-(CH₂)₂—O-2′-bridge, 2′-LNA or other bicyclic or “bridged” nucleoside analog, and 2′-O-(N-methylcarbamate) or those comprising base analogs. In connection with 2′-modified nucleotides as described for the present disclosure, by “amino” is meant 2′-NH₂ or 2′-O—NH₂, which can be modified or unmodified. Such modified groups are described, e.g., in Eckstein et al., U.S. Pat. No. 5,672,695 and Matulic-Adamic et al., U.S. Pat. No. 6,248,878. “Modified nucleotides” of the instant invention can also include nucleotide analogs as described above.

In reference to the nucleic acid molecules of the present disclosure, modifications may exist upon these agents in patterns on one or both strands of the double stranded ribonucleic acid (dsRNA). As used herein, “alternating positions” refers to a pattern where every other nucleotide is a modified nucleotide or there is an unmodified nucleotide (e.g., an unmodified ribonucleotide) between every modified nucleotide over a defined length of a strand of the dsRNA (e.g., 5′-MNMNMN-3′; 3′-MNMNMN-5′; where M is a modified nucleotide and N is an unmodified nucleotide). The modification pattern starts from the first nucleotide position at either the 5′ or 3′ terminus according to a position numbering convention, e.g., as described herein (in certain embodiments, position 1 is designated in reference to the terminal residue of a strand following a projected Dicer cleavage event of a DsiRNA agent of the invention; thus, position 1 does not always constitute a 3′ terminal or 5′ terminal residue of a pre-processed agent of the invention). The pattern of modified nucleotides at alternating positions may run the full length of the strand, but in certain embodiments includes at least 4, 6, 8, 10, 12, 14 nucleotides containing at least 2, 3, 4, 5, 6 or 7 modified nucleotides, respectively. As used herein, “alternating pairs of positions” refers to a pattern where two consecutive modified nucleotides are separated by two consecutive unmodified nucleotides over a defined length of a strand of the dsRNA (e.g., 5′-MMNNMMNNMMNN-3′; 3′-MMNNMMNNMMNN-5′; where M is a modified nucleotide and N is an unmodified nucleotide). The modification pattern starts from the first nucleotide position at either the 5′ or 3′ terminus according to a position numbering convention such as those described herein. The pattern of modified nucleotides at alternating positions may run the full length of the strand, but preferably includes at least 8, 12, 16, 20, 24, 28 nucleotides containing at least 4, 6, 8, 10, 12 or 14 modified nucleotides, respectively. It is emphasized that the above modification patterns are exemplary and are not intended as limitations on the scope of the invention.

As used herein, “base analog” refers to a heterocyclic moiety which is located at the 1′ position of a nucleotide sugar moiety in a modified nucleotide that can be incorporated into a nucleic acid duplex (or the equivalent position in a nucleotide sugar moiety substitution that can be incorporated into a nucleic acid duplex). In the dsRNAs of the invention, a base analog is generally either a purine or pyrimidine base excluding the common bases guanine (G), cytosine (C), adenine (A), thymine (T), and uracil (U). Base analogs can duplex with other bases or base analogs in dsRNAs. Base analogs include those useful in the compounds and methods of the invention, e.g., those disclosed in U.S. Pat. Nos. 5,432,272 and 6,001,983 to Benner and US Patent Publication No. 20080213891 to Manoharan, which are herein incorporated by reference. Non-limiting examples of bases include hypoxanthine (I), xanthine (X), 33-D-ribofuranosyl-(2,6-diaminopyrimidine) (K), 3-β-D-ribofuranosyl-(1-methyl-pyrazolo[4,3-d]pyrimidine-5,7(4H,6H)-dione) (P), iso-cytosine (iso-C), iso-guanine (iso-G), 1-β-D-ribofuranosyl-(5-nitroindole), 1-β-D-ribofuranosyl-(3-nitropyrrole), 5-bromouracil, 2-aminopurine, 4-thio-dT, 7-(2-thienyl)-imidazo[4,5-b]pyridine (Ds) and pyrrole-2-carbaldehyde (Pa), 2-amino-6-(2-thienyl)purine (S), 2-oxopyridine (Y), difluorotolyl, 4-fluoro-6-methylbenzimidazole, 4-methylbenzimidazole, 3-methyl isocarbostyrilyl, 5-methyl isocarbostyrilyl, and 3-methyl-7-propynyl isocarbostyrilyl, 7-azaindolyl, 6-methyl-7-azaindolyl, imidizopyridinyl, 9-methyl-imidizopyridinyl, pyrrolopyrizinyl, isocarbostyrilyl, 7-propynyl isocarbostyrilyl, propynyl-7-azaindolyl, 2,4,5-trimethylphenyl, 4-methylindolyl, 4,6-dimethylindolyl, phenyl, napthalenyl, anthracenyl, phenanthracenyl, pyrenyl, stilbenzyl, tetracenyl, pentacenyl, and structural derivates thereof (Schweitzer et al., J. Org. Chem., 59:7238-7242 (1994); Berger et al., Nucleic Acids Research, 28(15):2911-2914 (2000); Moran et al., J. Am. Chem. Soc., 119:2056-2057 (1997); Morales et al., J. Am. Chem. Soc., 121:2323-2324 (1999); Guckian et al., J. Am. Chem. Soc., 118:8182-8183 (1996); Morales et al., J. Am. Chem. Soc., 122(6):1001-1007 (2000); McMinn et al., J. Am. Chem. Soc., 121:11585-11586 (1999); Guckian et al., J. Org. Chem., 63:9652-9656 (1998); Moran et al., Proc. Natl. Acad. Sci., 94:10506-10511 (1997); Das et al., J. Chem. Soc., Perkin Trans., 1:197-206 (2002); Shibata et al., J. Chem. Soc., Perkin Trans., 1: 1605-1611 (2001); Wu et al., J. Am. Chem. Soc., 122(32):7621-7632 (2000); O'Neill et al., J. Org. Chem., 67:5869-5875 (2002); Chaudhuri et al., J. Am. Chem. Soc., 117:10434-10442 (1995); and U.S. Pat. No. 6,218,108). Base analogs may also be a universal base.

As used herein, “universal base” refers to a heterocyclic moiety located at the 1′ position of a nucleotide sugar moiety in a modified nucleotide, or the equivalent position in a nucleotide sugar moiety substitution, that, when present in a nucleic acid duplex, can be positioned opposite more than one type of base without altering the double helical structure (e.g., the structure of the phosphate backbone). Additionally, the universal base does not destroy the ability of the single stranded nucleic acid in which it resides to duplex to a target nucleic acid. The ability of a single stranded nucleic acid containing a universal base to duplex a target nucleic can be assayed by methods apparent to one in the art (e.g., UV absorbance, circular dichroism, gel shift, single stranded nuclease sensitivity, etc.). Additionally, conditions under which duplex formation is observed may be varied to determine duplex stability or formation, e.g., temperature, as melting temperature (Tm) correlates with the stability of nucleic acid duplexes. Compared to a reference single stranded nucleic acid that is exactly complementary to a target nucleic acid, the single stranded nucleic acid containing a universal base forms a duplex with the target nucleic acid that has a lower Tm than a duplex formed with the complementary nucleic acid. However, compared to a reference single stranded nucleic acid in which the universal base has been replaced with a base to generate a single mismatch, the single stranded nucleic acid containing the universal base forms a duplex with the target nucleic acid that has a higher Tm than a duplex formed with the nucleic acid having the mismatched base.

Some universal bases are capable of base pairing by forming hydrogen bonds between the universal base and all of the bases guanine (G), cytosine (C), adenine (A), thymine (T), and uracil (U) under base pair forming conditions. A universal base is not a base that forms a base pair with only one single complementary base. In a duplex, a universal base may form no hydrogen bonds, one hydrogen bond, or more than one hydrogen bond with each of G, C, A, T, and U opposite to it on the opposite strand of a duplex. Preferably, the universal bases does not interact with the base opposite to it on the opposite strand of a duplex. In a duplex, base pairing between a universal base occurs without altering the double helical structure of the phosphate backbone. A universal base may also interact with bases in adjacent nucleotides on the same nucleic acid strand by stacking interactions. Such stacking interactions stabilize the duplex, especially in situations where the universal base does not form any hydrogen bonds with the base positioned opposite to it on the opposite strand of the duplex. Non-limiting examples of universal-binding nucleotides include inosine, 1-β-D-ribofuranosyl-5-nitroindole, and/or 1-β-D-ribofuranosyl-3-nitropyrrole (US Pat. Appl. Publ. No. 20070254362 to Quay et al.; Van Aerschot et al., An acyclic 5-nitroindazole nucleoside analogue as ambiguous nucleoside. Nucleic Acids Res. 1995 Nov. 11; 23(21):4363-70; Loakes et al., 3-Nitropyrrole and 5-nitroindole as universal bases in primers for DNA sequencing and PCR. Nucleic Acids Res. 1995 Jul. 11; 23(13):2361-6; Loakes and Brown, 5-Nitroindole as an universal base analogue. Nucleic Acids Res. 1994 Oct. 11; 22(20):4039-43).

As used herein, “loop” refers to a structure formed by a single strand of a nucleic acid, in which complementary regions that flank a particular single stranded nucleotide region hybridize in a way that the single stranded nucleotide region between the complementary regions is excluded from duplex formation or Watson-Crick base pairing. A loop is a single stranded nucleotide region of any length. Examples of loops include the unpaired nucleotides present in such structures as hairpins, stem loops, or extended loops.

As used herein, “extended loop” in the context of a dsRNA refers to a single stranded loop and in addition 1, 2, 3, 4, 5, 6 or up to 20 base pairs or duplexes flanking the loop. In an extended loop, nucleotides that flank the loop on the 5′ side form a duplex with nucleotides that flank the loop on the 3′ side. An extended loop may form a hairpin or stem loop.

As used herein, “tetraloop” in the context of a dsRNA refers to a loop (a single stranded region) consisting of four nucleotides that forms a stable secondary structure that contributes to the stability of adjacent Watson-Crick hybridized nucleotides. Without being limited to theory, a tetraloop may stabilize an adjacent Watson-Crick base pair by stacking interactions. In addition, interactions among the four nucleotides in a tetraloop include but are not limited to non-Watson-Crick base pairing, stacking interactions, hydrogen bonding, and contact interactions (Cheong et al., Nature 1990 Aug. 16; 346(6285):680-2; Heus and Pardi, Science 1991 Jul. 12; 253(5016):191-4). A tetraloop confers an increase in the melting temperature (Tm) of an adjacent duplex that is higher than expected from a simple model loop sequence consisting of four random bases. For example, a tetraloop can confer a melting temperature of at least 55° C. in 10 mM NaHPO₄ to a hairpin comprising a duplex of at least 2 base pairs in length. A tetraloop may contain ribonucleotides, deoxyribonucleotides, modified nucleotides, and combinations thereof. Examples of RNA tetraloops include the UNCG family of tetraloops (e.g., UUCG), the GNRA family of tetraloops (e.g., GAAA), and the CUUG tetraloop. (Woese et al., Proc Natl Acad Sci USA. 1990 November; 87(21):8467-71; Antao et al., Nucleic Acids Res. 1991 Nov. 11; 19(21):5901-5). Examples of DNA tetraloops include the d(GNNA) family of tetraloops (e.g., d(GTTA), the d(GNRA)) family of tetraloops, the d(GNAB) family of tetraloops, the d(CNNG) family of tetraloops, the d(TNCG) family of tetraloops (e.g., d(TTCG)). (Nakano et al. Biochemistry, 41 (48), 14281-14292, 2002; SHINJI et al. Nippon Kagakkai Koen Yokoshu VOL. 78th; NO. 2; PAGE. 731 (2000).)

As used herein, the term “siRNA” refers to a double stranded nucleic acid in which each strand comprises RNA, RNA analog(s) or RNA and DNA. The siRNA comprises between 19 and 23 nucleotides or comprises 21 nucleotides. The siRNA typically has 2 bp overhangs on the 3′ ends of each strand such that the duplex region in the siRNA comprises 17-21 nucleotides, or 19 nucleotides. Typically, the antisense strand of the siRNA is sufficiently complementary with the target sequence of the α-1 antitrypsin gene/RNA.

Where a first sequence is referred to as “substantially complementary” with respect to a second sequence herein, the two sequences can be fully complementary, or they may form one or more, but generally not more than 4, 3 or 2 mismatched base pairs upon hybridization, while retaining the ability to hybridize under the conditions most relevant to their ultimate application. However, where two oligonucleotides are designed to form, upon hybridization, one or more single stranded overhangs, such overhangs shall not be regarded as mismatches with regard to the determination of complementarity. For example, a dsRNA comprising one oligonucleotide 21 nucleotides in length and another oligonucleotide 23 nucleotides in length, wherein the longer oligonucleotide comprises a sequence of 21 nucleotides that is fully complementary to the shorter oligonucleotide, may yet be referred to as “fully complementary” for the purposes of the invention.

The term “double-stranded RNA” or “dsRNA”, as used herein, refers to a complex of ribonucleic acid molecules, having a duplex structure comprising two anti-parallel and substantially complementary, as defined above, nucleic acid strands. The two strands forming the duplex structure may be different portions of one larger RNA molecule, or they may be separate RNA molecules. Where separate RNA molecules, such dsRNA are often referred to as siRNA (“short interfering RNA”) or DsiRNA (“Dicer substrate siRNAs”). Where the two strands are part of one larger molecule, and therefore are connected by an uninterrupted chain of nucleotides between the 3′-end of one strand and the 5′ end of the respective other strand forming the duplex structure, the connecting RNA chain is referred to as a “hairpin loop”, “short hairpin RNA” or “shRNA”. Where the two strands are connected covalently by means other than an uninterrupted chain of nucleotides between the 3′-end of one strand and the 5′end of the respective other strand forming the duplex structure, the connecting structure is referred to as a “linker”. The RNA strands may have the same or a different number of nucleotides. The maximum number of base pairs is the number of nucleotides in the shortest strand of the dsRNA minus any overhangs that are present in the duplex. In addition to the duplex structure, a dsRNA may comprise one or more nucleotide overhangs. In addition, as used herein, “dsRNA” may include chemical modifications to ribonucleotides, internucleoside linkages, end-groups, caps, and conjugated moieties, including substantial modifications at multiple nucleotides and including all types of modifications disclosed herein or known in the art. Any such modifications, as used in an siRNA- or DsiRNA-type molecule, are encompassed by “dsRNA” for the purposes of this specification and claims.

The phrase “duplex region” refers to the region in two complementary or substantially complementary oligonucleotides that form base pairs with one another, either by Watson-Crick base pairing or other manner that allows for a duplex between oligonucleotide strands that are complementary or substantially complementary. For example, an oligonucleotide strand having 21 nucleotide units can base pair with another oligonucleotide of 21 nucleotide units, yet only 19 bases on each strand are complementary or substantially complementary, such that the “duplex region” consists of 19 base pairs. The remaining base pairs may, for example, exist as 5′ and 3′ overhangs. Further, within the duplex region, 100% complementarity is not required; substantial complementarity is allowable within a duplex region. Substantial complementarity refers to complementarity between the strands such that they are capable of annealing under biological conditions. Techniques to empirically determine if two strands are capable of annealing under biological conditions are well know in the art. Alternatively, two strands can be synthesized and added together under biological conditions to determine if they anneal to one another.

By definition, “sufficiently complementary” (contrasted with, e.g., “100% complementary”) allows for one or more mismatches to exist between a dsRNA of the invention and the target RNA or cDNA sequence (e.g., α-1 antitrypsin mRNA), provided that the dsRNA possesses complementarity sufficient to trigger the destruction of the target RNA by the RNAi machinery (e.g., the RISC complex) or process. In certain embodiments, a “sufficiently complementary” dsRNA of the invention can harbor one, two, three or even four or more mismatches between the dsRNA sequence and the target RNA or cDNA sequence (e.g., in certain such embodiments, the antisense strand of the dsRNA harbors one, two, three, four, five or even six or more mismatches when aligned with the target RNA or cDNA sequence). Additional consideration of the preferred location of such mismatches within certain dsRNAs of the instant invention is considered in greater detail below.

As used herein “DsiRNAmm” refers to a DisRNA having a “mismatch tolerant region” containing one, two, three or four mismatched base pairs of the duplex formed by the sense and antisense strands of the DsiRNA, where such mismatches are positioned within the DsiRNA at a location(s) lying between (and thus not including) the two terminal base pairs of either end of the DsiRNA. The structure and mismatch positioning of exemplary forms of DsiRNAmm compositions are described in greater detail below.

Single-stranded nucleic acids that base pair over a number of bases are said to “hybridize.” Hybridization is typically determined under physiological or biologically relevant conditions (e.g., intracellular: pH 7.2, 140 mM potassium ion; extracellular pH 7.4, 145 mM sodium ion). Hybridization conditions generally contain a monovalent cation and biologically acceptable buffer and may or may not contain a divalent cation, complex anions, e.g. gluconate from potassium gluconate, uncharged species such as sucrose, and inert polymers to reduce the activity of water in the sample, e.g. PEG. Such conditions include conditions under which base pairs can form.

Hybridization is measured by the temperature required to dissociate single stranded nucleic acids forming a duplex, i.e., (the melting temperature; Tm). Hybridization conditions are also conditions under which base pairs can form. Various conditions of stringency can be used to determine hybridization (see, e.g., Wahl, G. M. and S. L. Berger (1987) Methods Enzymol. 152:399; Kimmel, A. R. (1987) Methods Enzymol. 152:507). Stringent temperature conditions will ordinarily include temperatures of at least about 30° C., more preferably of at least about 37° C., and most preferably of at least about 42° C. The hybridization temperature for hybrids anticipated to be less than 50 base pairs in length should be 5-10° C. less than the melting temperature (Tm) of the hybrid, where Tm is determined according to the following equations. For hybrids less than 18 base pairs in length, Tm(° C.)=2(# of A+T bases)+4(# of G+C bases). For hybrids between 18 and 49 base pairs in length, Tm(° C.)=81.5+16.6(log 10[Na+])+0.41 (% G+C)−(600/N), where N is the number of bases in the hybrid, and [Na+] is the concentration of sodium ions in the hybridization buffer ([Na+] for 1×SSC=0.165 M). For example, a hybridization determination buffer is shown in Table 1.

TABLE 1 To make 50 final conc. Vender Cat # Lot # m.w./Stock mL solution NaCl  100 mM Sigma S-5150 41K8934   5M    1 ml. KCl   80 mM Sigma P-9541 70K0002 74.55 0.298 g MgCl₂   8 mM Sigma M-1028 120K8933   1M  0.4 mL sucrose 2% w/v Fisher BP220- 907105 342.3    1 g 212 Tris-HCl   16 mM Fisher BP1757- 12419   1M  0.8 mL 500 NaH₂PO₄   1 mM Sigma S-3193 52H- 120.0 0.006 g 029515 EDTA 0.02 mM Sigma E-7889 110K89271 0.5M    2 μL H₂O Sigma W-4502 51K2359 to 50 mL pH = 7.0 adjust with at 20° C. HCI

Useful variations on hybridization conditions will be readily apparent to those skilled in the art. Hybridization techniques are well known to those skilled in the art and are described, for example, in Benton and Davis (Science 196:180, 1977); Grunstein and Hogness (Proc. Natl. Acad. Sci., USA 72:3961, 1975); Ausubel et al. (Current Protocols in Molecular Biology, Wiley Interscience, New York, 2001); Berger and Kimmel (Antisense to Molecular Cloning Techniques, 1987, Academic Press, New York); and Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York.

As used herein, “oligonucleotide strand” is a single stranded nucleic acid molecule. An oligonucleotide may comprise ribonucleotides, deoxyribonucleotides, modified nucleotides (e.g., nucleotides with 2′ modifications, synthetic base analogs, etc.) or combinations thereof. Such modified oligonucleotides can be preferred over native forms because of properties such as, for example, enhanced cellular uptake and increased stability in the presence of nucleases.

As used herein, the term “ribonucleotide” encompasses natural and synthetic, unmodified and modified ribonucleotides. Modifications include changes to the sugar moiety, to the base moiety and/or to the linkages between ribonucleotides in the oligonucleotide. As used herein, the term “ribonucleotide” specifically excludes a deoxyribonucleotide, which is a nucleotide possessing a single proton group at the 2′ ribose ring position.

As used herein, the term “deoxyribonucleotide” encompasses natural and synthetic, unmodified and modified deoxyribonucleotides. Modifications include changes to the sugar moiety, to the base moiety and/or to the linkages between deoxyribonucleotide in the oligonucleotide. As used herein, the term “deoxyribonucleotide” also includes a modified ribonucleotide that does not permit Dicer cleavage of a dsRNA agent, e.g., a 2′-O-methyl ribonucleotide, a phosphorothioate-modified ribonucleotide residue, etc., that does not permit Dicer cleavage to occur at a bond of such a residue.

As used herein, the term “PS-NA” refers to a phosphorothioate-modified nucleotide residue. The term “PS-NA” therefore encompasses both phosphorothioate-modified ribonucleotides (“PS-RNAs”) and phosphorothioate-modified deoxyribonucleotides (“PS-DNAs”).

As used herein, “Dicer” refers to an endoribonuclease in the RNase III family that cleaves a dsRNA or dsRNA-containing molecule, e.g., double-stranded RNA (dsRNA) or pre-microRNA (miRNA), into double-stranded nucleic acid fragments 19-25 nucleotides long, usually with a two-base overhang on the 3′ end. With respect to certain dsRNAs of the invention (e.g., “DsiRNAs”), the duplex formed by a dsRNA region of an agent of the invention is recognized by Dicer and is a Dicer substrate on at least one strand of the duplex. Dicer catalyzes the first step in the RNA interference pathway, which consequently results in the degradation of a target RNA. The protein sequence of human Dicer is provided at the NCBI database under accession number NP_085124, hereby incorporated by reference.

Dicer “cleavage” can be determined as follows (e.g., see Collingwood et al., Oligonucleotides 18:187-200 (2008)). In a Dicer cleavage assay, RNA duplexes (100 pmol) are incubated in 20 μL of 20 mM Tris pH 8.0, 200 mM NaCl, 2.5 mM MgCl2 with or without 1 unit of recombinant human Dicer (Stratagene, La Jolla, Calif.) at 37° C. for 18-24 hours. Samples are desalted using a Performa SR 96-well plate (Edge Biosystems, Gaithersburg, Md.). Electrospray-ionization liquid chromatography mass spectroscopy (ESI-LCMS) of duplex RNAs pre- and post-treatment with Dicer is done using an Oligo HTCS system (Novatia, Princeton, N.J.; Hail et al., 2004), which consists of a ThermoFinnigan TSQ7000, Xcalibur data system, ProMass data processing software and Paradigm MS4 HPLC (Michrom BioResources, Auburn, Calif.). In this assay, Dicer cleavage occurs where at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or even 100% of the Dicer substrate dsRNA, (i.e., 25-30 bp, dsRNA, preferably 26-30 bp dsRNA) is cleaved to a shorter dsRNA (e.g., 19-23 bp dsRNA, preferably, 21-23 bp dsRNA).

As used herein, “Dicer cleavage site” refers to the sites at which Dicer cleaves a dsRNA (e.g., the dsRNA region of a DsiRNA agent of the invention). Dicer contains two RNase III domains which typically cleave both the sense and antisense strands of a dsRNA. The average distance between the RNase III domains and the PAZ domain determines the length of the short double-stranded nucleic acid fragments it produces and this distance can vary (Macrae et al. (2006) Science 311: 195-8). As shown in FIG. 1, Dicer is projected to cleave certain double-stranded ribonucleic acids of the instant invention that possess an antisense strand having a 2 nucleotide 3′ overhang at a site between the 21^(st) and 22^(nd) nucleotides removed from the 3′ terminus of the antisense strand, and at a corresponding site between the 21^(st) and 22^(nd) nucleotides removed from the 5′ terminus of the sense strand. The projected and/or prevalent Dicer cleavage site(s) for dsRNA molecules distinct from those depicted in FIG. 1 may be similarly identified via art-recognized methods, including those described in Macrae et al. While the Dicer cleavage events depicted in FIG. 1 generate 21 nucleotide siRNAs, it is noted that Dicer cleavage of a dsRNA (e.g., DsiRNA) can result in generation of Dicer-processed siRNA lengths of 19 to 23 nucleotides in length. Indeed, in certain embodiments, a double-stranded DNA region may be included within a dsRNA for purpose of directing prevalent Dicer excision of a typically non-preferred 19mer or 20mer siRNA, rather than a 21mer.

In certain embodiments, dsRNAs of the invention are Dicer substrate siRNAs (“DsiRNAs”). DsiRNAs can possess certain advantages as compared to inhibitory nucleic acids that are not dicer substrates (“non-DsiRNAs”). Such advantages include, but are not limited to, enhanced duration of effect of a DsiRNA relative to a non-DsiRNA, as well as enhanced inhibitory activity of a DsiRNA as compared to a non-DsiRNA (e.g., a 19-23mer siRNA) when each inhibitory nucleic acid is suitably formulated and assessed for inhibitory activity in a mammalian cell at the same concentration (in this latter scenario, the DsiRNA would be identified as more potent than the non-DsiRNA). Detection of the enhanced potency of a DsiRNA relative to a non-DsiRNA is often most readily achieved at a formulated concentration (e.g., transfection concentration of the dsRNA) that results in the DsiRNA eliciting approximately 30-70% knockdown activity upon a target RNA (e.g., a mRNA). For active DsiRNAs, such levels of knockdown activity are most often achieved at in vitro mammalian cell DsiRNA transfection concentrations of 1 nM or less of as suitably formulated, and in certain instances are observed at DsiRNA transfection concentrations of 200 pM or less, 100 pM or less, 50 pM or less, 20 pM or less, 10 pM or less, 5 pM or less, or even 1 pM or less. Indeed, due to the variability among DsiRNAs of the precise concentration at which 30-70% knockdown of a target RNA is observed, construction of an IC₅₀ curve via assessment of the inhibitory activity of DsiRNAs and non-DsiRNAs across a range of effective concentrations is a preferred method for detecting the enhanced potency of a DsiRNA relative to a non-DsiRNA inhibitory agent.

As used herein, “overhang” refers to unpaired nucleotides, in the context of a duplex having one or more free ends at the 5′ terminus or 3′ terminus of a dsRNA. In certain embodiments, the overhang is a 3′ or 5′ overhang on the antisense strand or sense strand. In some embodiments, the overhang is a 3′ overhang having a length of between one and six nucleotides, optionally one to five, one to four, one to three, one to two, two to six, two to five, two to four, two to three, three to six, three to five, three to four, four to six, four to five, five to six nucleotides, or one, two, three, four, five or six nucleotides. “Blunt” or “blunt end” means that there are no unpaired nucleotides at that end of the dsRNA, i.e., no nucleotide overhang. For clarity, chemical caps or non-nucleotide chemical moieties conjugated to the 3′ end or 5′ end of an siRNA are not considered in determining whether an siRNA has an overhang or is blunt ended. In certain embodiments, the invention provides a dsRNA molecule for inhibiting the expression of the α-1 antitrypsin target gene in a cell or mammal, wherein the dsRNA comprises an antisense strand comprising a region of complementarity which is complementary to at least a part of an mRNA formed in the expression of the α-1 antitrypsin target gene, and wherein the region of complementarity is less than 35 nucleotides in length, optionally 19-24 nucleotides in length or 25-30 nucleotides in length, and wherein the dsRNA, upon contact with a cell expressing the α-1 antitrypsin target gene, inhibits the expression of the α-1 antitrypsin target gene by at least 10%, 25%, or 40%.

As used herein, the term “RNA processing” refers to processing activities performed by components of the siRNA, miRNA or RNase H pathways (e.g., Drosha, Dicer, Argonaute2 or other RISC endoribonucleases, and RNaseH), which are described in greater detail below (see “RNA Processing” section below). The term is explicitly distinguished from the post-transcriptional processes of 5′ capping of RNA and degradation of RNA via non-RISC- or non-RNase H-mediated processes. Such “degradation” of an RNA can take several forms, e.g. deadenylation (removal of a 3′ poly(A) tail), and/or nuclease digestion of part or all of the body of the RNA by one or more of several endo- or exo-nucleases (e.g., RNase III, RNase P, RNase T1, RNase A (1, 2, 3, 4/5), oligonucleotidase, etc.).

By “homologous sequence” is meant a nucleotide sequence that is shared by one or more polynucleotide sequences, such as genes, gene transcripts and/or non-coding polynucleotides. For example, a homologous sequence can be a nucleotide sequence that is shared by two or more genes encoding related but different proteins, such as different members of a gene family, different protein epitopes, different protein isoforms or completely divergent genes, such as a cytokine and its corresponding receptors. A homologous sequence can be a nucleotide sequence that is shared by two or more non-coding polynucleotides, such as noncoding DNA or RNA, regulatory sequences, introns, and sites of transcriptional control or regulation. Homologous sequences can also include conserved sequence regions shared by more than one polynucleotide sequence. Homology does not need to be perfect homology (e.g., 100%), as partially homologous sequences are also contemplated by the instant invention (e.g., 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80% etc.). Indeed, design and use of the dsRNA agents of the instant invention contemplates the possibility of using such dsRNA agents not only against target RNAs of α-1 antitrypsin possessing perfect complementarity with the presently described dsRNA agents, but also against target α-1 antitrypsin RNAs possessing sequences that are, e.g., only 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80% etc. complementary to said dsRNA agents. Similarly, it is contemplated that the presently described dsRNA agents of the instant invention might be readily altered by the skilled artisan to enhance the extent of complementarity between said dsRNA agents and a target α-1 antitrypsin RNA, e.g., of a specific allelic variant of α-1 antitrypsin (e.g., an allele of enhanced therapeutic interest). Indeed, dsRNA agent sequences with insertions, deletions, and single point mutations relative to the target α-1 antitrypsin sequence can also be effective for inhibition. Alternatively, dsRNA agent sequences with nucleotide analog substitutions or insertions can be effective for inhibition.

Sequence identity may be determined by sequence comparison and alignment algorithms known in the art. To determine the percent identity of two nucleic acid sequences (or of two amino acid sequences), the sequences are aligned for comparison purposes (e.g., gaps can be introduced in the first sequence or second sequence for optimal alignment). The nucleotides (or amino acid residues) at corresponding nucleotide (or amino acid) positions are then compared. When a position in the first sequence is occupied by the same residue as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % homology=# of identical positions/total # of positions×100), optionally penalizing the score for the number of gaps introduced and/or length of gaps introduced.

The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. In one embodiment, the alignment generated over a certain portion of the sequence aligned having sufficient identity but not over portions having low degree of identity (i.e., a local alignment). A preferred, non-limiting example of a local alignment algorithm utilized for the comparison of sequences is the algorithm of Karlin and Altschul (1990) Proc. Natl. Acad. Sci. USA 87:2264-68, modified as in Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-77. Such an algorithm is incorporated into the BLAST programs (version 2.0) of Altschul, et al. (1990) J. Mol. Biol. 215:403-10.

In another embodiment, a gapped alignment, the alignment is optimized by introducing appropriate gaps, and percent identity is determined over the length of the aligned sequences (i.e., a gapped alignment). To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., (1997) Nucleic Acids Res. 25(17):3389-3402. In another embodiment, a global alignment the alignment is optimized by introducing appropriate gaps, and percent identity is determined over the entire length of the sequences aligned. (i.e., a global alignment). A preferred, non-limiting example of a mathematical algorithm utilized for the global comparison of sequences is the algorithm of Myers and Miller, CABIOS (1989). Such an algorithm is incorporated into the ALIGN program (version 2.0) which is part of the GCG sequence alignment software package. When utilizing the ALIGN program for comparing amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used.

Greater than 80% sequence identity, e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or even 100% sequence identity, between the dsRNA antisense strand and the portion of the α-1 antitrypsin RNA sequence is preferred. Alternatively, the dsRNA may be defined functionally as a nucleotide sequence (or oligonucleotide sequence) that is capable of hybridizing with a portion of the α-1 antitrypsin RNA (e.g., 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mM EDTA, 50° C. or 70° C. hybridization for 12-16 hours; followed by washing). Additional preferred hybridization conditions include hybridization at 70° C. in 1×SSC or 50° C. in 1×SSC, 50% formamide followed by washing at 70° C. in 0.3×SSC or hybridization at 70° C. in 4×SSC or 50° C. in 4×SSC, 50% formamide followed by washing at 67° C. in 1×SSC. The hybridization temperature for hybrids anticipated to be less than 50 base pairs in length should be 5-10° C. less than the melting temperature (Tm) of the hybrid, where Tm is determined according to the following equations. For hybrids less than 18 base pairs in length, Tm(° C.)=2(# of A+T bases)+4(# of G+C bases). For hybrids between 18 and 49 base pairs in length, Tm(° C.)=81.5+16.6(log 10[Na+])+0.41 (% G+C)-(600/N), where N is the number of bases in the hybrid, and [Na+] is the concentration of sodium ions in the hybridization buffer ([Na+] for 1×SSC=0.165 M). Additional examples of stringency conditions for polynucleotide hybridization are provided in Sambrook, J., E. F. Fritsch, and T. Maniatis, 1989, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., chapters 9 and 11, and Current Protocols in Molecular Biology, 1995, F. M. Ausubel et al., eds., John Wiley & Sons, Inc., sections 2.10 and 6.3-6.4. The length of the identical nucleotide sequences may be at least 10, 12, 15, 17, 20, 22, 25, 27 or 30 bases.

By “conserved sequence region” is meant, a nucleotide sequence of one or more regions in a polynucleotide does not vary significantly between generations or from one biological system, subject, or organism to another biological system, subject, or organism. The polynucleotide can include both coding and non-coding DNA and RNA.

By “sense region” is meant a nucleotide sequence of a dsRNA molecule having complementarity to an antisense region of the dsRNA molecule. In addition, the sense region of a dsRNA molecule can comprise a nucleic acid sequence having homology with a target nucleic acid sequence.

By “antisense region” is meant a nucleotide sequence of a dsRNA molecule having complementarity to a target nucleic acid sequence. In addition, the antisense region of a dsRNA molecule comprises a nucleic acid sequence having complementarity to a sense region of the dsRNA molecule.

As used herein, “antisense strand” refers to a single stranded nucleic acid molecule which has a sequence complementary to that of a target RNA. When the antisense strand contains modified nucleotides with base analogs, it is not necessarily complementary over its entire length, but must at least hybridize with a target RNA.

As used herein, “sense strand” refers to a single stranded nucleic acid molecule which has a sequence complementary to that of an antisense strand. When the antisense strand contains modified nucleotides with base analogs, the sense strand need not be complementary over the entire length of the antisense strand, but must at least duplex with the antisense strand.

As used herein, “guide strand” refers to a single stranded nucleic acid molecule of a dsRNA or dsRNA-containing molecule, which has a sequence sufficiently complementary to that of a target RNA to result in RNA interference. After cleavage of the dsRNA or dsRNA-containing molecule by Dicer, a fragment of the guide strand remains associated with RISC, binds a target RNA as a component of the RISC complex, and promotes cleavage of a target RNA by RISC. As used herein, the guide strand does not necessarily refer to a continuous single stranded nucleic acid and may comprise a discontinuity, preferably at a site that is cleaved by Dicer. A guide strand is an antisense strand.

As used herein, “passenger strand” refers to an oligonucleotide strand of a dsRNA or dsRNA-containing molecule, which has a sequence that is complementary to that of the guide strand. As used herein, the passenger strand does not necessarily refer to a continuous single stranded nucleic acid and may comprise a discontinuity, preferably at a site that is cleaved by Dicer. A passenger strand is a sense strand.

By “target nucleic acid” is meant a nucleic acid sequence whose expression, level or activity is to be modulated. The target nucleic acid can be DNA or RNA. For agents that target α-1 antitrypsin, in certain embodiments, the target nucleic acid is α-1 antitrypsin RNA, e.g., in certain embodiments, α-1 antitrypsin mRNA. α-1 antitrypsin RNA target sites can also interchangeably be referenced by corresponding cDNA sequences. Levels of α-1 antitrypsin may also be targeted via targeting of upstream effectors of α-1 antitrypsin, or the effects of modulated or misregulated α-1 antitrypsin may also be modulated by targeting of molecules downstream of α-1 antitrypsin in the α-1 antitrypsin signalling pathway.

By “complementarity” is meant that a nucleic acid can form hydrogen bond(s) with another nucleic acid sequence by either traditional Watson-Crick or other non-traditional types. In reference to the nucleic molecules of the present invention, the binding free energy for a nucleic acid molecule with its complementary sequence is sufficient to allow the relevant function of the nucleic acid to proceed, e.g., RNAi activity. Determination of binding free energies for nucleic acid molecules is well known in the art (see, e.g., Turner et al., 1987, CSH Symp. Quant. Biol. LII pp. 123-133; Frier et al., 1986, Proc. Nat. Acad. Sci. USA 83:9373-9377; Turner et al., 1987, J. Am. Chem. Soc. 109:3783-3785). A percent complementarity indicates the percentage of contiguous residues in a nucleic acid molecule that can form hydrogen bonds (e.g., Watson-Crick base pairing) with a second nucleic acid sequence (e.g., 5, 6, 7, 8, 9, or 10 nucleotides out of a total of 10 nucleotides in the first oligonucleotide being based paired to a second nucleic acid sequence having 10 nucleotides represents 50%, 60%, 70%, 80%, 90%, and 100% complementary respectively). “Perfectly complementary” means that all the contiguous residues of a nucleic acid sequence will hydrogen bond with the same number of contiguous residues in a second nucleic acid sequence. In one embodiment, a dsRNA molecule of the invention comprises 19 to 30 (e.g., 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more) nucleotides that are complementary to one or more target nucleic acid molecules or a portion thereof.

As used herein, a dsRNA, e.g., DsiRNA or siRNA, having a sequence “sufficiently complementary” to a target RNA or cDNA sequence (e.g., α-1 antitrypsin mRNA) means that the dsRNA has a sequence sufficient to trigger the destruction of the target RNA (where a cDNA sequence is recited, the RNA sequence corresponding to the recited cDNA sequence) by the RNAi machinery (e.g., the RISC complex) or process. For example, a dsRNA that is “sufficiently complementary” to a target RNA or cDNA sequence to trigger the destruction of the target RNA by the RNAi machinery or process can be identified as a dsRNA that causes a detectable reduction in the level of the target RNA in an appropriate assay of dsRNA activity (e.g., an in vitro assay as described in Example 2 below), or, in further examples, a dsRNA that is sufficiently complementary to a target RNA or cDNA sequence to trigger the destruction of the target RNA by the RNAi machinery or process can be identified as a dsRNA that produces at least a 5%, at least a 10%, at least a 15%, at least a 20%, at least a 25%, at least a 30%, at least a 35%, at least a 40%, at least a 45%, at least a 50%, at least a 55%, at least a 60%, at least a 65%, at least a 70%, at least a 75%, at least a 80%, at least a 85%, at least a 90%, at least a 95%, at least a 98% or at least a 99% reduction in the level of the target RNA in an appropriate assay of dsRNA activity. In additional examples, a dsRNA that is sufficiently complementary to a target RNA or cDNA sequence to trigger the destruction of the target RNA by the RNAi machinery or process can be identified based upon assessment of the duration of a certain level of inhibitory activity with respect to the target RNA or protein levels in a cell or organism. For example, a dsRNA that is sufficiently complementary to a target RNA or cDNA sequence to trigger the destruction of the target RNA by the RNAi machinery or process can be identified as a dsRNA capable of reducing target mRNA levels by at least 20% at least 48 hours post-administration of said dsRNA to a cell or organism. Preferably, a dsRNA that is sufficiently complementary to a target RNA or cDNA sequence to trigger the destruction of the target RNA by the RNAi machinery or process is identified as a dsRNA capable of reducing target mRNA levels by at least 40% at least 72 hours post-administration of said dsRNA to a cell or organism, by at least 40% at least four, five or seven days post-administration of said dsRNA to a cell or organism, by at least 50% at least 48 hours post-administration of said dsRNA to a cell or organism, by at least 50% at least 72 hours post-administration of said dsRNA to a cell or organism, by at least 50% at least four, five or seven days post-administration of said dsRNA to a cell or organism, by at least 80% at least 48 hours post-administration of said dsRNA to a cell or organism, by at least 80% at least 72 hours post-administration of said dsRNA to a cell or organism, or by at least 80% at least four, five or seven days post-administration of said dsRNA to a cell or organism.

In certain embodiments, a nucleic acid of the invention (e.g., a DsiRNA or siRNA) possesses a sequence “sufficiently complementary to hybridize” to a target RNA or cDNA sequence, thereby achieving an inhibitory effect upon the target RNA. Hybridization, and conditions available for determining whether one nucleic acid is sufficiently complementary to another nucleic acid to allow the two sequences to hybridize, is described in greater detail below.

As used herein “cell” is used in its usual biological sense, and does not refer to an entire multicellular organism, e.g., specifically does not refer to a human. The cell can be present in an organism, e.g., birds, plants and mammals such as humans, cows, sheep, apes, monkeys, swine, dogs, and cats. The cell can be prokaryotic (e.g., bacterial cell) or eukaryotic (e.g., mammalian or plant cell). The cell can be of somatic or germ line origin, totipotent or pluripotent, dividing or non-dividing. The cell can also be derived from or can comprise a gamete or embryo, a stem cell, or a fully differentiated cell. Within certain aspects, the term “cell” refers specifically to mammalian cells, such as human cells, that contain one or more dsRNA molecules of the present disclosure. In particular aspects, a cell processes dsRNAs or dsRNA-containing molecules resulting in RNA interference of target nucleic acids, and contains proteins and protein complexes required for RNAi, e.g., Dicer and RISC.

By “RNA” is meant a molecule comprising at least one, and preferably at least 4, 8 and 12 ribonucleotide residues. The at least 4, 8 or 12 RNA residues may be contiguous. By “ribonucleotide” is meant a nucleotide with a hydroxyl group at the 2′ position of a β-D-ribofuranose moiety. The terms include double-stranded RNA, single-stranded RNA, RNA such as partially purified RNA, essentially pure RNA, synthetic RNA, recombinantly produced RNA, as well as altered RNA that differs from naturally occurring RNA by the addition, deletion, substitution and/or alteration of one or more nucleotides. Such alterations can include addition of non-nucleotide material, such as to the end(s) of the dsRNA or internally, for example at one or more nucleotides of the RNA. Nucleotides in the RNA molecules of the instant invention can also comprise non-standard nucleotides, such as non-naturally occurring nucleotides or chemically synthesized nucleotides or deoxynucleotides. These altered RNAs can be referred to as analogs or analogs of naturally-occurring RNA.

In certain embodiments, an RNAi agent (e.g., dsRNA) of the invention can be an “isolated” RNAi agent, meaning that the RNAi agent is isolated from (removed and/or purified from) a natural environment.

In some embodiments, an RNAi agent (e.g., dsRNA) of the invention can be a “synthetic” RNAi agent. The term “synthetic” or “non-natural” refers to an RNAi agent (e.g., a dsRNA of the disclosure) that (i) is synthesized using a machine or (ii) that is not derived from a cell or organism that normally produces the RNAi agent.

By “subject” is meant an organism, which is a donor or recipient of explanted cells or the cells themselves. “Subject” also refers to an organism to which the dsRNA agents of the invention can be administered. A subject can be a mammal or mammalian cells, including a human or human cells.

The phrase “pharmaceutically acceptable carrier” refers to a carrier for the administration of a therapeutic agent. Exemplary carriers include saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations thereof. For drugs administered orally, pharmaceutically acceptable carriers include, but are not limited to pharmaceutically acceptable excipients such as inert diluents, disintegrating agents, binding agents, lubricating agents, sweetening agents, flavoring agents, coloring agents and preservatives. Suitable inert diluents include sodium and calcium carbonate, sodium and calcium phosphate, and lactose, while corn starch and alginic acid are suitable disintegrating agents. Binding agents may include starch and gelatin, while the lubricating agent, if present, will generally be magnesium stearate, stearic acid or talc. If desired, the tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate, to delay absorption in the gastrointestinal tract. The pharmaceutically acceptable carrier of the disclosed dsRNA compositions may be micellar structures, such as a liposomes, capsids, capsoids, polymeric nanocapsules, or polymeric microcapsules.

Polymeric nanocapsules or microcapsules facilitate transport and release of the encapsulated or bound dsRNA into the cell. They include polymeric and monomeric materials, especially including polybutylcyanoacrylate. A summary of materials and fabrication methods has been published (see Kreuter, 1991). The polymeric materials which are formed from monomeric and/or oligomeric precursors in the polymerization/nanoparticle generation step, are per se known from the prior art, as are the molecular weights and molecular weight distribution of the polymeric material which a person skilled in the field of manufacturing nanoparticles may suitably select in accordance with the usual skill.

The term “in vitro” has its art recognized meaning, e.g., involving purified reagents or extracts, e.g., cell extracts. The term “in vivo” also has its art recognized meaning, e.g., involving living cells, e.g., immortalized cells, primary cells, cell lines, and/or cells in an organism.

“Treatment”, or “treating” as used herein, is defined as the application or administration of a therapeutic agent (e.g., a dsRNA agent or a vector or transgene encoding same) to a patient, or application or administration of a therapeutic agent to an isolated tissue or cell line from a patient, who has a disorder with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve or affect the disease or disorder, or symptoms of the disease or disorder. The term “treatment” or “treating” is also used herein in the context of administering agents prophylactically. The term “effective dose” or “effective dosage” is defined as an amount sufficient to achieve or at least partially achieve the desired effect. The term “therapeutically effective dose” is defined as an amount sufficient to cure or at least partially arrest the disease and its complications in a patient already suffering from the disease. The term “patient” includes human and other mammalian subjects that receive either prophylactic or therapeutic treatment.

Various methodologies of the instant invention include at least one step that involves comparing a value, level, feature, characteristic, property, etc. to a “suitable control”, referred to interchangeably herein as an “appropriate control”. A “suitable control” or “appropriate control” is a control or standard familiar to one of ordinary skill in the art useful for comparison purposes. In one embodiment, a “suitable control” or “appropriate control” is a value, level, feature, characteristic, property, etc. determined prior to performing an RNAi methodology, as described herein. For example, a transcription rate, mRNA level, translation rate, protein level, biological activity, cellular characteristic or property, genotype, phenotype, etc. can be determined prior to introducing an RNA silencing agent (e.g., DsiRNA) of the invention into a cell or organism. In another embodiment, a “suitable control” or “appropriate control” is a value, level, feature, characteristic, property, etc. determined in a cell or organism, e.g., a control or normal cell or organism, exhibiting, for example, normal traits. In yet another embodiment, a “suitable control” or “appropriate control” is a predefined value, level, feature, characteristic, property, etc.

α-1 Antitrypsin as an RNAi Target

Alpha 1-antitrypsin (AAT or Serpina1) is a protease inhibitor belonging to the serpin superfamily. It is generally known as serum trypsin inhibitor. Alpha 1-antitrypsin is also referred to as alpha-1 proteinase inhibitor (A1PI) because it inhibits a wide variety of proteases (Gettins PG. Chem Rev 102: 4751-804). It protects tissues from enzymes of inflammatory cells, especially neutrophil elastase, and has a reference range in blood of 1.5-3.5 gram/liter, but multi-fold elevated levels can occur upon acute inflammation (Kushner, Mackiewicz. Acute-phase glycoproteins: molecular biology, biochemistry and clinical applications (CRC Press). pp. 3-19). In the absence of AAT, neutrophil elastase is free to break down elastin, which contributes to the elasticity of the lungs, resulting in respiratory complications such as emphysema, or COPD (chronic obstructive pulmonary disease) in adults and cirrhosis in adults or children. Individuals with mutations in one or both copies of the AAT gene can suffer from alpha-1 anti-trypsin deficiency, which presents as a risk of developing pulmonary emphysema or chronic liver disease due to greater than normal elastase activity in the lungs and liver.

As mentioned above, in certain disease states associated with α-1 antitrypsin expression, an individual is producing significant quantities of alpha-1 antitrypsin, but a significant proportion of the α-1 antitrypsin protein being produced is misfolded or contains mutations that compromise the functioning of the protein. In certain such cases, the individual is producing misfolded proteins which cannot be properly transported from the site of synthesis to the site of action within the body.

Liver disease resulting from α-1 antitrypsin deficiency can be caused by such misfolded proteins. Mutant forms of α-1 antitrypsin (e.g., the common PiZ variant, which harbors a glutamate to lysine mutation at position 342 (position 366 in pre-processed form)) are produced in liver cells (hepatocytes in the liver commonly produce a large amount of circulating AAT), and in the misfolded configuration, such forms are not readily transported out of the cells. This leads to a buildup of misfolded protein in the liver cells and can cause one or more diseases or disorders of the liver including, but not limited to, chronic liver disease, liver inflammation, cirrhosis, liver fibrosis, and/or hepatocellular carcinoma.

RNAi therapies are newly identified to provide an attractive, targeted means of treating the effects of mutant forms of α-1 antitrypsin at a molecular level. Notably, RNAi therapies, such as the dsRNAs that are specifically exemplified herein, have demonstrated particularly good ability to be delivered to the cells of the liver in vivo (via, e.g., lipid nanoparticles and/or conjugates such as dynamic polyconjugates or GalNAc conjugates). Thus, formulated RNAi therapies, such as those described herein, are attractive modalities for treating or preventing diseases or disorders (especially the liver-specific diseases or disorders, e.g., chronic liver disease, liver inflammation, cirrhosis, liver fibrosis, and/or hepatocellular carcinoma) associated with mutant forms of α-1 antitrypsin.

Use of RNAi therapies to target α-1 antitrypsin at a molecular level in other tissues is also contemplated. Most notably, in the lung, the presence of mutant, inactive forms of α-1 antitrypsin in the serum can cause a deficiency in serum levels of active α-1 antitrypsin, resulting in host tissues that are susceptible to damage by neutrophil proteases. As a result, such lung tissues are rendered especially at risk of smoking injury, and lung-directed RNAi therapeutics such as those described herein offer an attractive and precise therapy.

α-1 Antitrypsin cDNA and Polypeptide Sequences

Known human and mouse α-1 antitrypsin cDNA and polypeptide sequences include the following: human α-1 antitrypsin NM_000295.4 and corresponding human α-1 antitrypsin polypeptide sequence GenBank Accession No. NP_000286.3; and mouse wild-type α-1 antitrypsin sequence GenBank Accession No. NM_009246.3 (Mus musculus C57BL/6 Serpina1d) and corresponding mouse Serpina1d sequence GenBank Accession No. NP_033272.1.

Assessment of α-1 Antitrypsin Levels

In certain embodiments, dsRNA-mediated inhibition of a α-1 antitrypsin target sequence is assessed. In such embodiments, α-1 antitrypsin RNA levels can be assessed by art-recognized methods (e.g., RT-PCR, Northern blot, expression array, etc.), optionally via comparison of α-1 antitrypsin levels in the presence of an anti-α-1 antitrypsin dsRNA of the invention relative to the absence of such an anti-α-1 antitrypsin dsRNA. In certain embodiments, α-1 antitrypsin levels in the presence of an anti-α-1 antitrypsin dsRNA are compared to those observed in the presence of vehicle alone, in the presence of a dsRNA directed against an unrelated target RNA, or in the absence of any treatment.

It is also recognized that levels of α-1 antitrypsin protein can be assessed and that α-1 antitrypsin protein levels are, under different conditions, either directly or indirectly related to α-1 antitrypsin RNA levels and/or the extent to which a dsRNA inhibits α-1 antitrypsin expression, thus art-recognized methods of assessing α-1 antitrypsin protein levels (e.g., Western blot, immunoprecipitation, other antibody-based methods, etc.) can also be employed to examine the inhibitory effect of a dsRNA of the invention.

In certain embodiments, potency of a dsRNA of the invention is determined in reference to the number of copies of a dsRNA present in the cytoplasm of a target cell that are required to achieve a certain level of target gene knockdown. For example, in certain embodiments, a potent dsRNA is one capable of causing 50% or greater knockdown of a target mRNA when present in the cytoplasm of a target cell at a copy number of 1000 or fewer RISC-loaded antisense strands per cell. More preferably, a potent dsRNA is one capable of producing 50% or greater knockdown of a target mRNA when present in the cytoplasm of a target cell at a copy number of 500 or fewer RISC-loaded antisense strands per cell. Optionally, a potent dsRNA is one capable of producing 50% or greater knockdown of a target mRNA when present in the cytoplasm of a target cell at a copy number of 300 or fewer RISC-loaded antisense strands per cell.

In further embodiments, the potency of a DsiRNA of the invention can be defined in reference to a 19 to 23mer dsRNA directed to the same target sequence within the same target gene. For example, a DsiRNA of the invention that possesses enhanced potency relative to a corresponding 19 to 23mer dsRNA can be a DsiRNA that reduces a target gene by an additional 5% or more, an additional 10% or more, an additional 20% or more, an additional 30% or more, an additional 40% or more, or an additional 50% or more as compared to a corresponding 19 to 23mer dsRNA, when assayed in an in vitro assay as described herein at a sufficiently low concentration to allow for detection of a potency difference (e.g., transfection concentrations at or below 1 nM in the environment of a cell, at or below 100 pM in the environment of a cell, at or below 10 pM in the environment of a cell, at or below 1 nM in the environment of a cell, in an in vitro assay as described herein; notably, it is recognized that potency differences can be best detected via performance of such assays across a range of concentrations—e.g., 0.1 pM to 10 nM—for purpose of generating a dose-response curve and identifying an IC₅₀ value associated with a DsiRNA/dsRNA).

In certain embodiments, a nucleic acid of the invention is administered in an amount sufficient to reduce α-1 antitrypsin target mRNA expression when the nucleic acid is introduced into a mammalian cell. In exemplary embodiments, reduction of α-1 antitrypsin target mRNA expression is assessed to have occurred if α-1 antitrypsin target mRNA levels are decreased by at least 5% or more, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more, as compared to a corresponding mammalian cell administered an appropriate control that does not include the α-1 antitryp sin-targeting nucleic acid of the invention. In an exemplary embodiment, the mammalian cell used to determine whether reduction of α-1 antitrypsin target mRNA expression has occurred relative to an appropriate control is a Huh7 cell, and the α-1 antitrypsin-targeting nucleic acid of the invention is optionally administered via transfection (e.g., using Lipofectamine™) at a concentration of at or below 1 nM in the environment of a cell, at or below 100 pM in the environment of a cell, at or below 10 pM in the environment of a cell, at or below 1 nM in the environment of a cell, in an in vitro assay as described herein.

α-1 antitrypsin inhibitory levels and/or α-1 antitrypsin levels may also be assessed indirectly, e.g., measurement of a reduction of the size, number and/or rate of growth or spread of polyps or tumors in a subject may be used to assess α-1 antitrypsin levels and/or α-1 antitrypsin inhibitory efficacy of a double-stranded nucleic acid of the instant invention.

Models Useful to Evaluate the Down-Regulation of α-1 Antitrypsin mRNA Levels and Expression

Therapeutic agents can be tested in selected animal model(s). For example, a dsRNA agent (or expression vector or transgene encoding same) as described herein can be used in an animal model to determine the efficacy, toxicity, or side effects of treatment with said agent. Alternatively, an agent (e.g., a therapeutic agent) can be used in an animal model to determine the mechanism of action of such an agent.

Cell Culture

The dsRNA agents of the invention can be tested for cleavage activity in vivo, for example, using the following procedure. The nucleotide sequences within the α-1 antitrypsin cDNA targeted by the dsRNA agents of the invention are shown in the above α-1 antitrypsin sequences.

The dsRNA reagents of the invention can be tested in cell culture using Huh7 or other mammalian cells (e.g., human cell lines Hep3B, HepG2, DU145, Calu3, SW480, T84, PL45, HeLa etc., and mouse cell lines AML12, Neuro2a, etc.) to determine the extent of α-1 antitrypsin RNA and α-1 antitrypsin protein inhibition. In certain embodiments, DsiRNA reagents (e.g., see FIG. 1, and exemplary structures recited elsewhere herein) are selected against the α-1 antitrypsin target as described herein. α-1 antitrypsin RNA inhibition is measured after delivery of these reagents by a suitable transfection agent to, for example, cultured Huh7 cells or other transformed or non-transformed mammalian cells in culture. Relative amounts of target α-1 antitrypsin RNA are measured versus HPRT1, actin or other appropriate control using real-time PCR monitoring of amplification (e.g., ABI 7700 TAQMAN®). A comparison is made to the activity of oligonucleotide sequences made to unrelated targets or to a randomized DsiRNA control with the same overall length and chemistry, or simply to appropriate vehicle-treated or untreated controls. Primary and secondary lead reagents are chosen for the target and optimization performed.

TAQMAN® (Real-Time PCR Monitoring of Amplification) and Lightcycler Quantification of mRNA

Total RNA is prepared from cells following DsiRNA delivery, for example, using Ambion Rnaqueous 4-PCR purification kit for large scale extractions, or Promega SV96 for 96-well assays. For Taqman analysis, dual-labeled probes are synthesized with, for example, the reporter dyes FAM or VIC covalently linked at the 5′-end and the quencher dye TAMRA conjugated to the 3′-end. PCR amplifications are performed on, for example, an ABI PRISM 7700 Sequence detector using 50 uL reactions consisting of 10 uL total RNA, 100 nM forward primer, 100 mM reverse primer, 100 nM probe, 1×TaqMan PCR reaction buffer (PE-Applied Biosystems), 5.5 mM MgCl2, 100 uM each dATP, dCTP, dGTP and dTTP, 0.2U RNase Inhibitor (Promega), 0.025U AmpliTaq Gold (PE-Applied Biosystems) and 0.2U M-MLV Reverse Transcriptase (Promega). The thermal cycling conditions can consist of 30 minutes at 48° C., 10 minutes at 95° C., followed by 40 cycles of 15 seconds at 95° C. and 1 minute at 60° C. Quantitation of target α-1 antitrypsin mRNA level is determined relative to standards generated from serially diluted total cellular RNA (300, 100, 30, 10 ng/rxn) and normalizing to, for example, HPRT1 mRNA in either parallel or same tube TaqMan reactions.

Western Blotting

Cellular protein extracts can be prepared using a standard micro preparation technique (for example using RIPA buffer), or preferably, by extracting nuclear proteins by a method such as the NE-PER Nuclear and Cytoplasmic Extraction kit (Thermo-Fisher Scientific). Cellular protein extracts are run on Tris-Glycine polyacrylamide gel and transferred onto membranes. Non-specific binding can be blocked by incubation, for example, with 5% non-fat milk for 1 hour followed by primary antibody for 16 hours at 4° C. Following washes, the secondary antibody is applied, for example (1:10,000 dilution) for 1 hour at room temperature and the signal detected on a VersaDoc imaging system

In several cell culture systems, cationic lipids have been shown to enhance the bioavailability of oligonucleotides to cells in culture (Bennet, et al., 1992, Mol. Pharmacology, 41, 1023-1033). In one embodiment, dsRNA molecules of the invention are complexed with cationic lipids for cell culture experiments. dsRNA and cationic lipid mixtures are prepared in serum-free OptimMEM (InVitrogen) immediately prior to addition to the cells. OptiMEM is warmed to room temperature (about 20-25° C.) and cationic lipid is added to the final desired concentration. dsRNA molecules are added to OptiMEM to the desired concentration and the solution is added to the diluted dsRNA and incubated for 15 minutes at room temperature. In dose response experiments, the RNA complex is serially diluted into OptiMEM prior to addition of the cationic lipid.

Animal Models

The efficacy of anti-α-1 antitrypsin dsRNA agents may be evaluated in an animal model. Animal models of liver diseases, conditions, or disorders as are known in the art can be used for evaluation of the efficacy, potency, toxicity, etc. of anti-α-1 antitrypsin dsRNAs. Exemplary animals model of α-1 antitrypsin-induced liver disease are transgenic mice that express human α-1 antitrypsin mutant Z protein, PiZ, which recapitulate the human liver disease and exhibit inflammation, increased levels of apoptosis and autophagy, accelerated proliferation and enhanced development of hepatic progenitor cells (the behavior of such models also suggest the employment of antioxidants in combination with anti-α-1 antitrypsin dsRNAs in treatment of liver disease; Marcus et al. Exper Biol Med 237: 1163-1172; Teckman et al. Am J Physiol Gastrointest Liver Physiol 286: G851-62; Rudnick et al. Hepatology 39: 1048-55; Brunt et al. J Pediatr Gastroenterol Nutr 51: 626-30). Such mice are available, for example, from Saint Louis University. These animal models may be used as a source cells or tissue for assays of the compositions of the invention. Such models can also be used or adapted for use for pre-clinical evaluation of the efficacy of dsRNA compositions of the invention in modulating α-1 antitrypsin gene expression toward therapeutic use.

Such models and/or wild-type mice can be used in evaluating the efficacy of dsRNA molecules of the invention to inhibit α-1 antitrypsin levels, expression, development of α-1 antitrypsin-associated phenotypes, diseases or disorders, etc. These models, wild-type mice and/or other models can similarly be used to evaluate the safety/toxicity and efficacy of dsRNA molecules of the invention in a pre-clinical setting.

Specific examples of animal model systems useful for evaluation of the α-1 antitrypsin-targeting dsRNAs of the invention include wild-type mice and transgenic PiZ mutant model mice. In an exemplary in vivo experiment, dsRNAs of the invention are tail vein injected into such mouse models at doses ranging from 1 to 10 mg/kg or, alternatively, repeated doses are administered at single-dose IC₅₀ levels, and organ samples (e.g., liver, but may also include prostate, kidney, lung, pancreas, colon, skin, spleen, bone marrow, lymph nodes, mammary fat pad, etc.) are harvested 24 hours after administration of the final dose. Such organs are then evaluated for mouse and/or human α-1 antitrypsin levels, depending upon the model used. Duration of action can also be examined at, e.g., 1, 4, 7, 14, 21 or more days after final dsRNA administration.

α-1 Antitrypsin-Targeting dsRNAs

In certain embodiments, an anti-α-1 antitrypsin DsiRNA of the instant invention possesses strand lengths of at least 25 nucleotides. Accordingly, in certain embodiments, an anti-α-1 antitrypsin DsiRNA contains one oligonucleotide sequence, a first sequence, that is at least 25 nucleotides in length and no longer than 35 or up to 50 or more nucleotides. This sequence of RNA can be between 26 and 35, 26 and 34, 26 and 33, 26 and 32, 26 and 31, 26 and 30, and 26 and 29 nucleotides in length. This sequence can be 27 or 28 nucleotides in length or 27 nucleotides in length. The second sequence of the DsiRNA agent can be a sequence that anneals to the first sequence under biological conditions, such as within the cytoplasm of a eukaryotic cell. Generally, the second oligonucleotide sequence will have at least 19 complementary base pairs with the first oligonucleotide sequence, more typically the second oligonucleotide sequence will have 21 or more complementary base pairs, or 25 or more complementary base pairs with the first oligonucleotide sequence. In one embodiment, the second sequence is the same length as the first sequence, and the DsiRNA agent is blunt ended. In another embodiment, the ends of the DsiRNA agent have one or more overhangs.

In certain embodiments, the first and second oligonucleotide sequences of the DsiRNA agent exist on separate oligonucleotide strands that can be and typically are chemically synthesized. In some embodiments, both strands are between 26 and 35 nucleotides in length. In other embodiments, both strands are between 25 and 30 or 26 and 30 nucleotides in length. In one embodiment, both strands are 27 nucleotides in length, are completely complementary and have blunt ends. In certain embodiments of the instant invention, the first and second sequences of an anti-α-1 antitrypsin DsiRNA exist on separate RNA oligonucleotides (strands). In one embodiment, one or both oligonucleotide strands are capable of serving as a substrate for Dicer. In other embodiments, at least one modification is present that promotes Dicer to bind to the double-stranded RNA structure in an orientation that maximizes the double-stranded RNA structure's effectiveness in inhibiting gene expression. In certain embodiments of the instant invention, the anti-α-1 antitrypsin DsiRNA agent is comprised of two oligonucleotide strands of differing lengths, with the anti-α-1 antitrypsin DsiRNA possessing a blunt end at the 3′ terminus of a first strand (sense strand) and a 3′ overhang at the 3′ terminus of a second strand (antisense strand). The DsiRNA can also contain one or more deoxyribonucleic acid (DNA) base substitutions.

Suitable DsiRNA compositions that contain two separate oligonucleotides can be chemically linked outside their annealing region by chemical linking groups. Many suitable chemical linking groups are known in the art and can be used. Suitable groups will not block Dicer activity on the DsiRNA and will not interfere with the directed destruction of the RNA transcribed from the target gene. Alternatively, the two separate oligonucleotides can be linked by a third oligonucleotide such that a hairpin structure is produced upon annealing of the two oligonucleotides making up the DsiRNA composition. The hairpin structure will not block Dicer activity on the DsiRNA and will not interfere with the directed destruction of the target RNA.

The dsRNA molecule can be designed such that every residue of the antisense strand is complementary to a residue in the target molecule. Alternatively, substitutions can be made within the molecule to increase stability and/or enhance processing activity of said molecule. Substitutions can be made within the strand or can be made to residues at the ends of the strand. In certain embodiments, substitutions and/or modifications are made at specific residues within a DsiRNA agent. Such substitutions and/or modifications can include, e.g., deoxy-modifications at one or more residues of positions 1, 2 and 3 when numbering from the 3′ terminal position of the sense strand of a DsiRNA agent; and introduction of 2′-O-alkyl (e.g., 2′-O-methyl) modifications at the 3′ terminal residue of the antisense strand of DsiRNA agents, with such modifications also being performed at overhang positions of the 3′ portion of the antisense strand and at alternating residues of the antisense strand of the DsiRNA that are included within the region of a DsiRNA agent that is processed to form an active siRNA agent. The preceding modifications are offered as exemplary, and are not intended to be limiting in any manner. Further consideration of the structure of preferred DsiRNA agents, including further description of the modifications and substitutions that can be performed upon the anti-α-1 antitrypsin DsiRNA agents of the instant invention, can be found below.

A dsRNA of the invention comprises two RNA strands that are sufficiently complementary to hybridize to form a duplex structure. One strand of the dsRNA (the antisense strand) comprises a region of complementarity that is substantially complementary, and generally fully complementary, to a target sequence, derived from the sequence of an mRNA formed during the expression of the α-1 antitrypsin target gene, the other strand (the sense strand) comprises a region which is complementary to the antisense strand, such that the two strands hybridize and form a duplex structure when combined under suitable conditions. Generally, the duplex structure is between 15 and 35, optionally between 25 and 30, between 26 and 30, between 18 and 25, between 19 and 24, or between 19 and 21 base pairs in length. Similarly, the region of complementarity to the target sequence is between 15 and 35, optionally between 18 and 30, between 25 and 30, between 19 and 24, or between 19 and 21 nucleotides in length. The dsRNA of the invention may further comprise one or more single-stranded nucleotide overhang(s). It has been identified that dsRNAs comprising duplex structures of between 15 and 35 base pairs in length can be effective in inducing RNA interference, including DsiRNAs (generally of at least 25 base pairs in length) and siRNAs (in certain embodiments, duplex structures of siRNAs are between 20 and 23, and optionally, specifically 21 base pairs (Elbashir et al., EMBO 20: 6877-6888)). It has also been identified that dsRNAs possessing duplexes shorter than 20 base pairs can be effective as well (e.g., 15, 16, 17, 18 or 19 base pair duplexes). In certain embodiments, the dsRNAs of the invention can comprise at least one strand of a length of 19 nucleotides or more. In certain embodiments, it can be reasonably expected that shorter dsRNAs comprising a sequence complementary to one of the sequences of Tables 5, 10, 15 or 20, minus only a few nucleotides on one or both ends may be similarly effective as compared to the dsRNAs described above and in Tables 2-3, 7-8, 12-13 and 17-18. Hence, dsRNAs comprising a partial sequence of at least 15, 16, 17, 18, 19, 20, or more contiguous nucleotides sufficiently complementary to one of the sequences of Tables 5, 10, 15 or 20, and differing in their ability to inhibit the expression of the α-1 antitrypsin target gene in an assay as described herein by not more than 5, 10, 15, 20, 25, or 30% inhibition from a dsRNA comprising the full sequence, are contemplated by the invention. In one embodiment, at least one end of the dsRNA has a single-stranded nucleotide overhang of 1 to 5, optionally 1 to 4, in certain embodiments, 1 or 2 nucleotides. Certain dsRNA structures having at least one nucleotide overhang possess superior inhibitory properties as compared to counterparts possessing base-paired blunt ends at both ends of the dsRNA molecule.

In one embodiment, dsRNA molecules of the invention that down regulate or reduce α-1 antitrypsin gene expression are used for treating, preventing or reducing α-1 antitrypsin-related diseases or disorders (e.g., liver disease) in a subject or organism.

In one embodiment of the present invention, each sequence of a DsiRNA molecule of the invention is independently 25 to 35 nucleotides in length, in specific embodiments 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35 nucleotides in length. In another embodiment, the DsiRNA duplexes of the invention independently comprise 25 to 30 base pairs (e.g., 25, 26, 27, 28, 29, or 30). In another embodiment, one or more strands of the DsiRNA molecule of the invention independently comprises 19 to 35 nucleotides (e.g., 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35) that are complementary to a target (α-1 antitrypsin) nucleic acid molecule. In certain embodiments, a DsiRNA molecule of the invention possesses a length of duplexed nucleotides between 25 and 34 nucleotides in length (e.g., 25, 26, 27, 28, 29, 30, 31, 32, 33 or 34 nucleotides in length; optionally, all such nucleotides base pair with cognate nucleotides of the opposite strand). (Exemplary DsiRNA molecules of the invention are shown in FIG. 1, and below).

In certain additional embodiments of the present invention, each oligonucleotide of a DsiRNA molecule of the invention is independently 25 to 53 nucleotides in length, in specific embodiments 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52 or 53 nucleotides in length. For DsiRNAs possessing a strand that exceeds 30 nucleotides in length, available structures include those where only one strand exceeds 30 nucleotides in length (see, e.g., U.S. Pat. No. 8,349,809), or those where both strands exceed 30 nucleotides in length (see, e.g., WO 2010/080129). Stabilizing modifications (e.g., 2′-O-Methyl, phosphorothioate, deoxyribonucleotides, including dNTP base pairs, etc.) can be incorporated within any double stranded nucleic acid of the invention, and can be used in particular within DsiRNAs possessing one or both strands exceeding 30 nucleotides in length. While the guide strand of a double stranded nucleic acid of the invention must possess a sequence of, e.g., 15, 16, 17, 18 or 19 nucleotides that are complementary to a target RNA (e.g., mRNA), additional sequence(s) of the guide strand need not be complementary to the target RNA. The end structures of double stranded nucleic acids possessing at least one strand length in excess of 30 nucleotides can also be varied while continuing to yield functional dsNAs—e.g. the 5′ end of the guide strand and the 3′ end of the passenger strand may form a 5′-overhang, a blunt end or a 3′ overhang (for certain dsNAs, e.g., “single strand extended” dsNAs, the length of such a 5′ or 3′ overhang can be 1-4, 1-5, 1-6, 1-10, 1-15, 1-20 or even 1-25 or more); similarly, the 3′ end of the guide strand and the 5′ end of the passenger strand may form a 5′-overhang, a blunt end or a 3′ overhang (for certain dsNAs, e.g., “single strand extended” dsNAs, the length of such a 5′ or 3′ overhang can be 1-4, 1-5, 1-6, 1-10, 1-15, 1-20 or even 1-25 or more). In certain embodiments, the length of the passenger strand is 31-49 nucleotides while the length of the guide strand is 31-53 nucleotides, optionally while the 5′ end of the guide strand forms a blunt end (optionally, a base-paired blunt end) with the 3′ end of the passenger strand, optionally, with the 3′ end of the guide strand and the 5′ end of the passenger strand forming a 3′ overhang of 1-4 nucleotides in length. Exemplary “extended” Dicer substrate structures are set forth, e.g., in US 2010/0173974 and U.S. Pat. No. 8,349,809, both of which are incorporated herein by reference. In certain embodiments, one or more strands of the dsNA molecule of the invention independently comprises 19 to 35 nucleotides (e.g., 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35) that are complementary to a target (α-1 antitrypsin) nucleic acid molecule. In certain embodiments, a DsiRNA molecule of the invention possesses a length of duplexed nucleotides between 25 and 49 nucleotides in length (e.g., 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 or 49 nucleotides in length; optionally, all such nucleotides base pair with cognate nucleotides of the opposite strand).

In certain embodiments, a DsiRNA (in a state as initially formed, prior to dicer cleavage) is more potent at reducing α-1 antitrypsin target gene expression in a mammalian cell than a 19, 20, 21, 22 or 23 base pair sequence that is contained within it. In certain such embodiments, a DsiRNA prior to dicer cleavage is more potent than a 19-21mer contained within it. Optionally, a DsiRNA prior to dicer cleavage is more potent than a 19 base pair duplex contained within it that is synthesized with symmetric dTdT overhangs (thereby forming a siRNA possessing 21 nucleotide strand lengths having dTdT overhangs). In certain embodiments, the DsiRNA is more potent than a 19-23mer siRNA (e.g., a 19 base pair duplex with dTdT overhangs) that targets at least 19 nucleotides of the 21 nucleotide target sequence that is recited for a DsiRNA of the invention (without wishing to be bound by theory, the identity of a such a target site for a DsiRNA is identified via identification of the Ago2 cleavage site for the DsiRNA; once the Ago2 cleavage site of a DsiRNA is determined for a DsiRNA, identification of the Ago2 cleavage site for any other inhibitory dsRNA can be performed and these Ago2 cleavage sites can be aligned, thereby determining the alignment of projected target nucleotide sequences for multiple dsRNAs). In certain related embodiments, the DsiRNA is more potent than a 19-23mer siRNA that targets at least 20 nucleotides of the 21 nucleotide target sequence that is recited for a DsiRNA of the invention. Optionally, the DsiRNA is more potent than a 19-23mer siRNA that targets the same 21 nucleotide target sequence that is recited for a DsiRNA of the invention. In certain embodiments, the DsiRNA is more potent than any 21mer siRNA that targets the same 21 nucleotide target sequence that is recited for a DsiRNA of the invention. Optionally, the DsiRNA is more potent than any 21 or 22mer siRNA that targets the same 21 nucleotide target sequence that is recited for a DsiRNA of the invention. In certain embodiments, the DsiRNA is more potent than any 21, 22 or 23mer siRNA that targets the same 21 nucleotide target sequence that is recited for a DsiRNA of the invention. As noted above, such potency assessments are most effectively performed upon dsRNAs that are suitably formulated (e.g., formulated with an appropriate transfection reagent) at a concentration of 1 nM or less. Optionally, an IC₅₀ assessment is performed to evaluate activity across a range of effective inhibitory concentrations, thereby allowing for robust comparison of the relative potencies of dsRNAs so assayed.

The dsRNA molecules of the invention are added directly, or can be complexed with lipids (e.g., cationic lipids), packaged within liposomes, or otherwise delivered to target cells or tissues. The nucleic acid or nucleic acid complexes can be locally administered to relevant tissues ex vivo, or in vivo through direct dermal application, transdermal application, or injection, with or without their incorporation in biopolymers. In particular embodiments, the nucleic acid molecules of the invention comprise sequences shown in FIG. 1, and the below exemplary structures. Examples of such nucleic acid molecules consist essentially of sequences defined in these figures and exemplary structures. Furthermore, where such agents are modified in accordance with the below description of modification patterning of DsiRNA agents, chemically modified forms of constructs described in FIG. 1, and the below exemplary structures can be used in all uses described for the DsiRNA agents of FIG. 1, and the below exemplary structures.

In another aspect, the invention provides mammalian cells containing one or more dsRNA molecules of this invention. The one or more dsRNA molecules can independently be targeted to the same or different sites.

Modified Structures of Anti-α-1 Antitrypsin DsiRNA Agents

In certain embodiments, the anti-α-1 antitrypsin DsiRNA agents of the invention can have the following structures:

In one such embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers. In a related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, and “D”=DNA. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand.

DsiRNAs of the invention can carry a broad range of modification patterns (e.g., 2′-O-methyl RNA patterns, e.g., within extended DsiRNA agents). Certain modification patterns of the second strand of DsiRNAs of the invention are presented below.

In one embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. In a related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand.

In another such embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. In a related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand.

In another such embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. In a related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. In a related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M7” or “M7” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. In a related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M6” or “M6” modification pattern.

In other embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In a related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M5” or “M5” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In a related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M4” or “M4” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In a related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M8” or “M8” modification pattern.

In other embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M3” or “M3” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In a related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M2” or “M2” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In a related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M1” or “M1” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M9” or “M9” modification pattern.

In other embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In a related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M10” or “M10” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M11” or “M11” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M12” or “M12” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M13” or “M13” modification pattern.

In other embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M21” or “M21” modification pattern.

In further embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M14” or “M14” modification pattern.

In additional embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M15” or “M15” modification pattern.

In further embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In a related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M16” or “M16” modification pattern.

In additional embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M17” or “M17” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M18” or “M18” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M19” or “M19” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M20” or “M20” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M22” or “M22” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M24” or “M24” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M25” or “M25” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M26” or “M26” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M27” or “M27” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M28” or “M28” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M29” or “M29” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M30” or “M30” modification pattern.

In further embodiments, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXXX-3' 3'-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXDD-3' 3'-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M31” or “M31” modification pattern.

In additional embodiments, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXXX-3' 3'-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXDD-3' 3'-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M32” or “M32” modification pattern.

In further embodiments, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXXX-3' 3'-YXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXDD-3' 3'-YXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXXX-3' 3'-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXDD-3' 3'-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M34” or “M34” modification pattern.

In additional embodiments, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXXX-3' 3'-YXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXDD-3' 3'-YXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXXX-3' 3'-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXDD-3' 3'-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M35” or “M35” modification pattern.

In further embodiments, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXXX-3' 3'-YXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXDD-3' 3'-YXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5'-XXXXXXXXXXXXXXXXXXXXXXXXX-3' 3'-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5' wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M37” or “M37” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M38” or “M38” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M40” or “M40” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M41” or “M41” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M36” or “M36” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M42” or “M42” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M43” or “M43” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M44” or “M44” modification pattern.

In further embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M45” or “M45” modification pattern.

In additional embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M46” or “M46” modification pattern.

In further embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M47” or “M47” modification pattern.

In additional embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M48” or “M48” modification pattern.

In further embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M52” or “M52” modification pattern.

In further embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M54” or “M54” modification pattern.

In additional embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M55” or “M55” modification pattern.

In further embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M56” or “M56” modification pattern.

In additional embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M57” or “M57” modification pattern.

In further embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M58” or “M58” modification pattern.

In additional embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M59” or “M59” modification pattern.

In further embodiments, the DsiRNA comprises:

 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M60” or “M60” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M61” or “M61” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M62” or “M62” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M63” or “M63” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M64” or “M64” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M65” or “M65” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M66” or “M66” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M67” or “M67” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M68” or “M68” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M69” or “M69” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M70” or “M70” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′  wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M71” or “M71” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M72” or “M72” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers and underlined residues are 2′-O-methyl RNA monomers. The top strand is the sense strand, and the bottom strand is the antisense strand. In one related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M73” or “M73” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M7*” or “M7*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M6*” or “M6*” modification pattern.

In other embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M5*” or “M5*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M4*” or “M4*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M8*” or “M8*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M2*” or “M2*” modification pattern.

In other embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M10*” or “M10*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M11*” or “M11*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M13*” or “M13*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M14*” or “M14*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M15*” or “M15*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M16*” or “M16*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M17*” or “M17*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M18*” or “M18*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M19*” or “M19*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, underlined residues are 2′-O-methyl RNA monomers, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In another related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M20*” or “M20*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M22*” or “M22*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M24*” or “M24*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M25*” or “M25*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M26*” or “M26*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M27*” or “M27*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M28*” or “M28*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M29*” or “M29*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M34*” or “M34*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M35*” or “M35*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M37*” or “M37*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M38*” or “M38*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M40*” or “M40*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M41*” or “M41*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M36*” or “M36*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M42*” or “M42*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M43*” or “M43*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M44*” or “M44*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M46*” or “M46*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M47*” or “M47*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M48*” or “M48*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M52*” or “M52*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M54*” or “M54*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M55*” or “M55*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M56*” or “M56*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M57*” or “M57*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M58*” or “M58*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M59*” or “M59*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M60*” or “M60*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M61*” or “M61*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M62*” or “M62*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M63*” or “M63*” modification pattern.

In further embodiments, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M64*” or “M64*” modification pattern.

In additional embodiments, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M65*” or “M65*” modification pattern.

In further embodiments, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M66*” or “M66*” modification pattern.

In additional embodiments, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M67*” or “M67*” modification pattern.

In further embodiments, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M68*” or “M68*” modification pattern.

In additional embodiments, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M69*” or “M69*” modification pattern.

In further embodiments, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M70*” or “M70*” modification pattern.

In additional embodiments, the DsiRNA comprises:

  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M71*” or “M71*” modification pattern.

In further embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M72*” or “M72*” modification pattern.

In additional embodiments, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA and “X”=2′-O-methyl RNA. The top strand is the sense strand, and the bottom strand is the antisense strand. In a further related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, and “D”=DNA. The top strand is the sense strand, and the bottom strand is the antisense strand. This modification pattern is also referred to herein as the “AS-M73*” or “M73*” modification pattern. Additional exemplary antisense strand modifications include the following:

3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M74” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M75” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M76” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M77” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M78” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M79” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M80” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M81” 3′-XpXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M82” 3′-XpXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M83” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M84” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M85” 3′-FFFXXXXXXXXXXXXXXXXXXXXFFFF-5′ “AS-M88” 3′-XXXXFXXXFXFXXXXXXXXXXXXXXXX-5′ “AS-M89” 3′-FFFXFXFXFXFXFXFXFXXXXXXFFFF-5′ “AS-M90” 3′-FFFXXXXXXXXXXXXXXXXXXXXXXFF-5′ “AS-M91” 3′-XXXXXXFXXXXXFXFXXXXXXXXXXXX-5′ “AS-M92” 3′-FFFXFXXXXXXXXXXXXXFXXXXXXXX-5′ “AS-M93” 3′-FFFXFXFXXXXXFXFXFXFXXXXFFFF-5′ “AS-M94” 3′-FFFXFXFXFXFXFXFXFXXXXXXFFFF-5′ “AS-M95” 3′-FFFXFXFXFXFXFXFXFXXXXXXFFFpF-5′ “AS-M96” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M210” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M74*” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M75*” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M76*” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M77*” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M78*” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M79*” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M80*” 3′-XpXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M82*” 3′-XpXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M83*” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M84*” 3′-XFFXXXXXXXXXXXXXXXXXXXXFFFF-5′ “AS-M88*” 3′-XXXXFXXXFXFXXXXXXXXXXXXXXXX-5′ “AS-M89*” 3′-XFFXFXFXFXFXFXFXFXXXXXXFFFF-5′ “AS-M90*” 3′-XFFXXXXXXXXXXXXXXXXXXXXXXFF-5′ “AS-M91*” 3′-XXXXXXFXXXXXFXFXXXXXXXXXXXX-5′ “AS-M92*” 3′-XFFXFXXXXXXXXXXXXXFXXXXXXXX-5′ “AS-M93*” 3′-XFFXFXFXXXXXFXFXFXFXXXXFFFF-5′ “AS-M94*” 3′-XFFXFXFXFXFXFXFXFXXXXXXFFFF-5′ “AS-M95*” 3′-XFFXFXFXFXFXFXFXFXXXXXXFFFpF-5′ “AS-M96*” 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ “AS-M210*” where “X”=RNA, “X”=2′-O-methyl RNA, “D”=DNA, “F”=2′-Fluoro NA and “p”=Phosphorothioate linkage. In certain additional embodiments, the antisense strand of selected dsRNAs of the invention are extended, optionally at the 5′ end, with an exemplary 5′ extension of base “AS-M8”, “AS-M17” and “AS-M48” modification patterns respectively represented as follows:

(SEQ ID NO: 3493) 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXXU

GCU

UCGT-5′ “AS-M8, extended” (SEQ ID NO: 3493) 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXXU

GCU

UCGT-5′ “AS-M17, extended” (SEQ ID NO: 3493) 3′-XXXXXXXXXXXXXXXXXXXXXXXXXXXU

GCU

UCGT-5′ “AS-M48, extended” where “X”=RNA; “X”=2′-O-methyl RNA; “F”=2′-Fluoro NA and “A” in bold, italics indicates a 2′-Fluoro-adenine residue.

In certain embodiments, the sense strand of a DsiRNA of the invention is modified—specific exemplary forms of sense strand modifications are shown below, and it is contemplated that such modified sense strands can be substituted for the sense strand of any of the DsiRNAs shown above to generate a DsiRNA comprising a below-depicted sense strand that anneals with an above-depicted antisense strand. Exemplary sense strand modification patterns include:

5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM1” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM2” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM3” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM4” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM5” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM6” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM7” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM8” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM9” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM10” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM11” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM12” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM13” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM14” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM15” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM16” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM17” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM18” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM19” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM20” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM21” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM23” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM24” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM25” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM30” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM31” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM32” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM33” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM34” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM35” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM36” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM37” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM38” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM39” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM40” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM41” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM42” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM43” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM44” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM45″, “SM47” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM46” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM48” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM49” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM50” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM51” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM52” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM53” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM54” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM55” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM56” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM57” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM58” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM59” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM60” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM61” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM62” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM63” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM64” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM65” 5′-XXXXXXXXXXXXXXXXXXXXXXXpDpD-3′ “SM66” 5′-XpXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM67” 5′-XpXXXXXXXXXXXXXXXXXXXXXXpDpD-3′ “SM68” 5′-DXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM69” 5′-DpXXXXXXXXXXXXXXXXXXXXXXpDpD-3′ “SM70” 5′-DXDXXXXXXXXXXXXXXXXXXXXDD-3′ “SM71” 5′-DpXDXXXXXXXXXXXXXXXXXXXXDD-3′ “SM72” 5′-XXDXXXXXXXXXDXXXXXXXXXXDD-3′ “SM73” 5′-XpXDXXXXXXXXXDXXXXXXXXXXDD-3′ “SM74” 5′-DXDXXXXXXXXXDXXXXXDXXXXDD-3′ “SM75” 5′-DpXDXXXXXXXXXDXXXXXDXXXXDD-3′ “SM76” 5′-XpXpXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM77” 5′-XpXpXXXXXXXXXXXXXXXXXXXXXpDpD-3′ “SM78” 5′-DpXpDXXXXXXXXXXXXXXXXXXXXDD-3′ “SM79” 5′-XpXpDXXXXXXXXXDXXXXXXXXXXDD-3′ “SM80” 5′-DXDXXXDXXXXXDXXXXXDXXXXDD-3′ “SM81” 5′-DpXDXXXDXXXXXDXXXXXDXXXXpDpD-3′ “SM82” 5′ C3 spacer-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM83” 5′ C3 spacer-XXDXXXXXXXXXDXXXXXXXXXXDD-3′ “SM84” 5′ C3 spacer-XXXXXXXXXXXXXXXXXXXXXXXpDpD-3′ “SM85” 5′-XXXXXXXXXXXXXXXXXXXXXXXpDpD-3′ “SM86” 5′-XpXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM87” 5′-XpXXXXXXXXXXXXXXXXXXXXXXpDpD-3′ “SM88” 5′-DXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM89” 5′-DpXXXXXXXXXXXXXXXXXXXXXXpDpD-3′ “SM90” 5′-DXDXXXXXXXXXXXXXXXXXXXXDD-3′ “SM91” 5′-DpXDXXXXXXXXXXXXXXXXXXXXDD-3′ “SM92” 5′-XXDXXXXXXXXXDXXXXXXXXXXDD-3′ “SM93” 5′-XpXDXXXXXXXXXDXXXXXXXXXXDD-3′ “SM94” 5′-DXDXXXXXXXXXDXXXXXDXXXXDD-3′ “SM95” 5′-DpXDXXXXXXXXXDXXXXXDXXXXDD-3′ “SM96” 5′-XpXpXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM97” 5′-XpXpXXXXXXXXXXXXXXXXXXXXXpDpD-3′ “SM98” 5′-DpXpDXXXXXXXXXXXXXXXXXXXXDD-3′ “SM99” 5′-XpXpDXXXXXXXXXDXXXXXXXXXXDD-3′ “SM100” 5′-DXDXXXXXXXXXDXDXXXDDXXDDD-3′ “SM101” 5′-DpXDXXXDXXXXXDXXXXXDXXXXpDpD-3′ “SM102” 5′ C3 spacer-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM103” 5′ C3 spacer-XXDXXXXXXXXXDXXXXXXXXXXDD-3′ “SM104” 5′ C3 spacer-XXXXXXXXXXXXXXXXXXXXXXXpDpD-3′ “SM105” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM106” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM107” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM108” 5′-XFXXXXXXXXXXXXXXXFXXXXXDD-3′ “SM110” 5′-XXXFXFXXXXXXXFXXXXXXXXXDD-3′ “SM111” 5′-XFXFXFXFXXXFXFXFXFXXXXXDD-3′ “SM112” 5′-XpFXFXFXFXXXFXFXFXFXXXXXpDpD-3′ “SM113” 5′-XFXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM114” 5′-XXXXFFXFXXXFXFXXXXXXXXXDD-3′ “SM115” 5′-XFXFXXXXXXXXXXXXFXXXXXXDD-3′ “SM116” 5′-XFXFFFXFXXXFXFXFFFXXXXXDD-3′ “SM117” 5′-XFXFXFXFXXXFXFXFXFXXXXXDD-3′ “SM118” 5′-XpFXFXFXFXXXFXFXFXFXXXXXpDpD-3′ “SM119” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM250” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM251” 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ “SM252” 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ “SM22” 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′  5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ 5′-XXXXXXXXXXXXXXXXXXXXXXXXX-3′ where “X”=RNA, “X”=2′-O-methyl RNA, “D”=DNA, “F”=2′-Fluoro NA and “p”=Phosphorothioate linkage.

The above modification patterns can also be incorporated into, e.g., the extended DsiRNA structures and mismatch and/or frayed DsiRNA structures described below.

In another embodiment, the DsiRNA comprises strands having equal lengths possessing 1-3 mismatched residues that serve to orient Dicer cleavage (specifically, one or more of positions 1, 2 or 3 on the first strand of the DsiRNA, when numbering from the 3′-terminal residue, are mismatched with corresponding residues of the 5′-terminal region on the second strand when first and second strands are annealed to one another). An exemplary 27mer DsiRNA agent with two terminal mismatched residues is shown:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX

3′-XXXXXXXXXXXXXXXXXXXXXXXXX

wherein “X”=RNA, “M”=Nucleic acid residues (RNA, DNA or non-natural or modified nucleic acids) that do not base pair (hydrogen bond) with corresponding “M” residues of otherwise complementary strand when strands are annealed. Any of the residues of such agents can optionally be 2′-O-methyl RNA monomers—alternating positioning of 2′-O-methyl RNA monomers that commences from the 3′-terminal residue of the bottom (second) strand, as shown for above asymmetric agents, can also be used in the above “blunt/fray” DsiRNA agent. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand.

In certain additional embodiments, the present invention provides compositions for RNA interference (RNAi) that possess one or more base paired deoxyribonucleotides within a region of a double stranded ribonucleic acid (dsRNA) that is positioned 3′ of a projected sense strand Dicer cleavage site and correspondingly 5′ of a projected antisense strand Dicer cleavage site. The compositions of the invention comprise a dsRNA which is a precursor molecule, i.e., the dsRNA of the present invention is processed in vivo to produce an active small interfering nucleic acid (siRNA). The dsRNA is processed by Dicer to an active siRNA which is incorporated into RISC.

In certain embodiments, the DsiRNA agents of the invention can have the following exemplary structures (noting that any of the following exemplary structures can be combined, e.g., with the bottom strand modification patterns of the above-described structures—in one specific example, the bottom strand modification pattern shown in any of the above structures is applied to the 27 most 3′ residues of the bottom strand of any of the following structures; in another specific example, the bottom strand modification pattern shown in any of the above structures upon the 23 most 3′ residues of the bottom strand is applied to the 23 most 3′ residues of the bottom strand of any of the following structures):

In one such embodiment, the DsiRNA comprises the following (an exemplary “right-extended”, “DNA extended” DsiRNA):

5′-XXXXXXXXXXXXXXXXXXXXXXXX_(N*)D_(N)DD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXX_(N*)D_(N)XX-5′ wherein “X”=RNA, “Y” is an optional overhang domain comprised of 0-10 RNA monomers that are optionally 2′-O-methyl RNA monomers—in certain embodiments, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, “D”=DNA, and “N”=1 to 50 or more, but is optionally 1-8 or 1-10. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand.

In a related embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXXX_(N*)D_(N)DD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXX_(N*)D_(N)DD-5′ wherein “X”=RNA, “Y” is an optional overhang domain comprised of 0-10 RNA monomers that are optionally 2′-O-methyl RNA monomers—in certain embodiments, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, “D”=DNA, and “N”=1 to 50 or more, but is optionally 1-8 or 1-10. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand.

In an additional embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXX_(N*)D_(N)DD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXX_(N*)D_(N)ZZ-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an optional overhang domain comprised of 0-10 RNA monomers that are optionally 2′-O-methyl RNA monomers—in certain embodiments, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, “D”=DNA, “Z”=DNA or RNA, and “N”=1 to 50 or more, but is optionally 1-8 or 1-10. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand, with 2′-O-methyl RNA monomers located at alternating residues along the top strand, rather than the bottom strand presently depicted in the above schematic.

In another such embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXX_(N*)D_(N)DD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXX_(N*)D_(N)ZZ-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an optional overhang domain comprised of 0-10 RNA monomers that are optionally 2′-O-methyl RNA monomers—in certain embodiments, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, “D”=DNA, “Z”=DNA or RNA, and “N”=1 to 50 or more, but is optionally 1-8 or 1-10. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand, with 2′-O-methyl RNA monomers located at alternating residues along the top strand, rather than the bottom strand presently depicted in the above schematic.

In another such embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXX_(N*)D_(N)DD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXX_(N*)D_(N)ZZ-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “Y” is an optional overhang domain comprised of 0-10 RNA monomers that are optionally 2′-O-methyl RNA monomers—in certain embodiments, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, “D”=DNA, “Z”=DNA or RNA, and “N”=1 to 50 or more, but is optionally 1-8 or 1-10. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand, with 2′-O-methyl RNA monomers located at alternating residues along the top strand, rather than the bottom strand presently depicted in the above schematic.

In another embodiment, the DsiRNA comprises:

5′-XXXXXXXXXXXXXXXXXXXXXXXX_(N*)[X1/D1]_(N)DD-3′ 3′-YXXXXXXXXXXXXXXXXXXXXXXXX_(N*)[X2/D2]_(N)ZZ-5′ wherein “X”=RNA, “Y” is an optional overhang domain comprised of 0-10 RNA monomers that are optionally 2′-O-methyl RNA monomers—in certain embodiments, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, “D”=DNA, “Z”=DNA or RNA, and “N”=1 to 50 or more, but is optionally 1-8 or 1-10, where at least one D1_(N) is present in the top strand and is base paired with a corresponding D2_(N) in the bottom strand. Optionally, D1_(N) and D1_(N+1) are base paired with corresponding D2_(N) and D2_(N+1); D1_(N), D1_(N+1) and D1_(N+2) are base paired with corresponding D2_(N), D1_(N+1) and D1_(N+2), etc. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand, with 2′-O-methyl RNA monomers located at alternating residues along the top strand, rather than the bottom strand presently depicted in the above schematic.

In the structures depicted herein, the 5′ end of either the sense strand or antisense strand can optionally comprise a phosphate group.

In another embodiment, a DNA:DNA-extended DsiRNA comprises strands having equal lengths possessing 1-3 mismatched residues that serve to orient Dicer cleavage (specifically, one or more of positions 1, 2 or 3 on the first strand of the DsiRNA, when numbering from the 3′-terminal residue, are mismatched with corresponding residues of the 5′-terminal region on the second strand when first and second strands are annealed to one another). An exemplary DNA:DNA-extended DsiRNA agent with two terminal mismatched residues is shown:

5′-D_(N)XXXXXXXXXXXXXXXXXXXXXXXXXX_(N*) ^(M) _(M-3′) 3′-D_(N)XXXXXXXXXXXXXXXXXXXXXXXXX

wherein “X”=RNA, “M”=Nucleic acid residues (RNA, DNA or non-natural or modified nucleic acids) that do not base pair (hydrogen bond) with corresponding “M” residues of otherwise complementary strand when strands are annealed, “D”=DNA and “N”=1 to 50 or more, but is optionally 1-15 or, optionally, 1-8. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. Any of the residues of such agents can optionally be 2′-O-methyl RNA monomers—alternating positioning of 2′-O-methyl RNA monomers that commences from the 3′-terminal residue of the bottom (second) strand, as shown for above asymmetric agents, can also be used in the above “blunt/fray” DsiRNA agent. In one embodiment, the top strand (first strand) is the sense strand, and the bottom strand (second strand) is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand. Modification and DNA:DNA extension patterns paralleling those shown above for asymmetric/overhang agents can also be incorporated into such “blunt/frayed” agents.

In one embodiment, a length-extended DsiRNA agent is provided that comprises deoxyribonucleotides positioned at sites modeled to function via specific direction of Dicer cleavage, yet which does not require the presence of a base-paired deoxyribonucleotide in the dsRNA structure. An exemplary structure for such a molecule is shown:

5′-XXXXXXXXXXXXXXXXXXXDDXX-3′ 3′-YXXXXXXXXXXXXXXXXXDDXXXX-5′ wherein “X”=RNA, “Y” is an optional overhang domain comprised of 0-10 RNA monomers that are optionally 2′-O-methyl RNA monomers—in certain embodiments, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, and “D”=DNA. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand. The above structure is modeled to force Dicer to cleave a minimum of a 21mer duplex as its primary post-processing form. In embodiments where the bottom strand of the above structure is the antisense strand, the positioning of two deoxyribonucleotide residues at the ultimate and penultimate residues of the 5′ end of the antisense strand will help reduce off-target effects (as prior studies have shown a 2′-O-methyl modification of at least the penultimate position from the 5′ terminus of the antisense strand to reduce off-target effects; see, e.g., US 2007/0223427).

In one embodiment, the DsiRNA comprises the following (an exemplary “left-extended”, “DNA extended” DsiRNA):

5′-D_(N)XXXXXXXXXXXXXXXXXXXXXXXX_(N)*Y-3′ 3′-D_(N)XXXXXXXXXXXXXXXXXXXXXXXX_(N)*-5′ wherein “X”=RNA, “Y” is an optional overhang domain comprised of 0-10 RNA monomers that are optionally 2′-O-methyl RNA monomers—in certain embodiments, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, “D”=DNA, and “N”=1 to 50 or more, but is optionally 1-8 or 1-10. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand.

In a related embodiment, the DsiRNA comprises:

5′-D_(N)XXXXXXXXXXXXXXXXXXXXXXXX_(N)*DD-3′ 3′-D_(N)XXXXXXXXXXXXXXXXXXXXXXXX_(N)*XX-5′ wherein “X”=RNA, optionally a 2′-O-methyl RNA monomers “D”=DNA, “N”=1 to 50 or more, but is optionally 1-8 or 1-10. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand.

In an additional embodiment, the DsiRNA comprises:

5′-D_(N)XXXXXXXXXXXXXXXXXXXXXXXX_(N)*DD-3′ 3′-D_(N) XXXXXXXXXXXXXXXXXXXXXXXX_(N)*ZZ-5′ wherein “X”=RNA, optionally a 2′-O-methyl RNA monomers “D”=DNA, “N”=1 to 50 or more, but is optionally 1-8 or 1-10. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. “Z”=DNA or RNA. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand, with 2′-O-methyl RNA monomers located at alternating residues along the top strand, rather than the bottom strand presently depicted in the above schematic.

In another such embodiment, the DsiRNA comprises:

5′-D_(N)XXXXXXXXXXXXXXXXXXXXXXXX_(N)*DD-3′ 3′-D_(N) XXXXXXXXXXXXXXXXXXXXXXXX_(N)*ZZ-5′ wherein “X”=RNA, optionally a 2′-O-methyl RNA monomers “D”=DNA, “N”=1 to 50 or more, but is optionally 1-8 or 1-10. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. “Z”=DNA or RNA. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand, with 2′-O-methyl RNA monomers located at alternating residues along the top strand, rather than the bottom strand presently depicted in the above schematic.

In another such embodiment, the DsiRNA comprises:

5′-D_(N)ZZXXXXXXXXXXXXXXXXXXXXXXXX_(N)*DD-3′ 3′-D_(N) XXXXXXXXXXXXXXXXXXXXXXXXXX_(N)*ZZ-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “D”=DNA, “Z”=DNA or RNA, and “N”=1 to 50 or more, but is optionally 1-8 or 1-10. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand, with 2′-O-methyl RNA monomers located at alternating residues along the top strand, rather than the bottom strand presently depicted in the above schematic.

In another such embodiment, the DsiRNA comprises:

5′-D_(N)ZZXXXXXXXXXXXXXXXXXXXXXXXX_(N)*Y-3′ 3′-D_(N) XXXXXXXXXXXXXXXXXXXXXXXXXX_(N)*-5′ wherein “X”=RNA, “X”=2′-O-methyl RNA, “D”=DNA, “Z”=DNA or RNA, and “N”=1 to 50 or more, but is optionally 1-8 or 1-10. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. “Y” is an optional overhang domain comprised of 0-10 RNA monomers that are optionally 2′-O-methyl RNA monomers—in certain embodiments, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand, with 2′-O-methyl RNA monomers located at alternating residues along the top strand, rather than the bottom strand presently depicted in the above schematic.

In another embodiment, the DsiRNA comprises:

5′-[X1/D1]_(N)XXXXXXXXXXXXXXXXXXXXXXXX_(N)*DD-3′ 3′-[X2/D2]_(N)XXXXXXXXXXXXXXXXXXXXXXXX_(N)*ZZ-5′ wherein “X”=RNA, “D”=DNA, “Z”=DNA or RNA, and “N”=1 to 50 or more, but is optionally 1-8 or 1-10, where at least one D1_(N) is present in the top strand and is base paired with a corresponding D2_(N) in the bottom strand. Optionally, D1_(N) and D1_(N+1) are base paired with corresponding D2_(N) and D2_(N+1); D1_(N), D1_(N+1) and D1_(N+2) are base paired with corresponding D2_(N), D1_(N+1) and D1_(N+2), etc. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand, with 2′-O-methyl RNA monomers located at alternating residues along the top strand, rather than the bottom strand presently depicted in the above schematic.

In a related embodiment, the DsiRNA comprises:

5′-[X1/D1]_(N)XXXXXXXXXXXXXXXXXXXXXXXX_(N)*Y-3′ 3′-[X2/D2]_(N)XXXXXXXXXXXXXXXXXXXXXXXX_(N)*-5′ wherein “X”=RNA, “D”=DNA, “Y” is an optional overhang domain comprised of 0-10 RNA monomers that are optionally 2′-O-methyl RNA monomers—in certain embodiments, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, and “N”=1 to 50 or more, but is optionally 1-8 or 1-10, where at least one D1_(N) is present in the top strand and is base paired with a corresponding D2_(N) in the bottom strand. Optionally, D1_(N) and D1_(N+1) are base paired with corresponding D2_(N) and D2_(N+1); D1_(N), D1_(N+1) and D1_(N+2) are base paired with corresponding D2_(N), D1_(N+1) and D1_(N+2), etc. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand, with 2′-O-methyl RNA monomers located at alternating residues along the top strand, rather than the bottom strand presently depicted in the above schematic.

In another embodiment, the DNA:DNA-extended DsiRNA comprises strands having equal lengths possessing 1-3 mismatched residues that serve to orient Dicer cleavage (specifically, one or more of positions 1, 2 or 3 on the first strand of the DsiRNA, when numbering from the 3′-terminal residue, are mismatched with corresponding residues of the 5′-terminal region on the second strand when first and second strands are annealed to one another). An exemplary DNA:DNA-extended DsiRNA agent with two terminal mismatched residues is shown:

5′-D_(N)XXXXXXXXXXXXXXXXXXXXXXXXXX_(N*) ^(M) _(M-3′) 3′-D_(N)XXXXXXXXXXXXXXXXXXXXXXXXXX

wherein “X”=RNA, “M”=Nucleic acid residues (RNA, DNA or non-natural or modified nucleic acids) that do not base pair (hydrogen bond) with corresponding “M” residues of otherwise complementary strand when strands are annealed, “D”=DNA and “N”=1 to 50 or more, but is optionally 1-8 or 1-10. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. Any of the residues of such agents can optionally be 2′-O-methyl RNA monomers—alternating positioning of 2′-O-methyl RNA monomers that commences from the 3′-terminal residue of the bottom (second) strand, as shown for above asymmetric agents, can also be used in the above “blunt/fray” DsiRNA agent. In one embodiment, the top strand (first strand) is the sense strand, and the bottom strand (second strand) is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand. Modification and DNA:DNA extension patterns paralleling those shown above for asymmetric/overhang agents can also be incorporated into such “blunt/frayed” agents.

In another embodiment, a length-extended DsiRNA agent is provided that comprises deoxyribonucleotides positioned at sites modeled to function via specific direction of Dicer cleavage, yet which does not require the presence of a base-paired deoxyribonucleotide in the dsRNA structure. Exemplary structures for such a molecule are shown:

5′-XXDDXXXXXXXXXXXXXXXXXXXX_(N)*Y-3′ 3′-DDXXXXXXXXXXXXXXXXXXXXXX_(N)*-5′ or

5′-XDXDXXXXXXXXXXXXXXXXXXXX_(N)*Y-3′ 3′-DXDXXXXXXXXXXXXXXXXXXXXX_(N)*-5′ wherein “X”=RNA, “Y” is an optional overhang domain comprised of 0-10 RNA monomers that are optionally 2′-O-methyl RNA monomers—in certain embodiments, “Y” is an overhang domain comprised of 1-4 RNA monomers that are optionally 2′-O-methyl RNA monomers, and “D”=DNA. “N*”=0 to 15 or more, but is optionally 0, 1, 2, 3, 4, 5 or 6. In one embodiment, the top strand is the sense strand, and the bottom strand is the antisense strand. Alternatively, the bottom strand is the sense strand and the top strand is the antisense strand.

In any of the above embodiments where the bottom strand of the above structure is the antisense strand, the positioning of two deoxyribonucleotide residues at the ultimate and penultimate residues of the 5′ end of the antisense strand will help reduce off-target effects (as prior studies have shown a 2′-O-methyl modification of at least the penultimate position from the 5′ terminus of the antisense strand to reduce off-target effects; see, e.g., US 2007/0223427).

In certain embodiments, the “D” residues of the above structures include at least one PS-DNA or PS-RNA. Optionally, the “D” residues of the above structures include at least one modified nucleotide that inhibits Dicer cleavage.

While the above-described “DNA-extended” DsiRNA agents can be categorized as either “left extended” or “right extended”, DsiRNA agents comprising both left- and right-extended DNA-containing sequences within a single agent (e.g., both flanks surrounding a core dsRNA structure are dsDNA extensions) can also be generated and used in similar manner to those described herein for “right-extended” and “left-extended” agents.

In some embodiments, the DsiRNA of the instant invention further comprises a linking moiety or domain that joins the sense and antisense strands of a DNA:DNA-extended DsiRNA agent. Optionally, such a linking moiety domain joins the 3′ end of the sense strand and the 5′ end of the antisense strand. The linking moiety may be a chemical (non-nucleotide) linker, such as an oligomethylenediol linker, oligoethylene glycol linker, or other art-recognized linker moiety. Alternatively, the linker can be a nucleotide linker, optionally including an extended loop and/or tetraloop.

In one embodiment, the DsiRNA agent has an asymmetric structure, with the sense strand having a 25-base pair length, and the antisense strand having a 27-base pair length with a 1-4 base 3′-overhang (e.g., a one base 3′-overhang, a two base 3′-overhang, a three base 3′-overhang or a four base 3′-overhang). In another embodiment, this DsiRNA agent has an asymmetric structure further containing 2 deoxynucleotides at the 3′ end of the sense strand.

In another embodiment, the DsiRNA agent has an asymmetric structure, with the antisense strand having a 25-base pair length, and the sense strand having a 27-base pair length with a 1-4 base 3′-overhang (e.g., a one base 3′-overhang, a two base 3′-overhang, a three base 3′-overhang or a four base 3′-overhang). In another embodiment, this DsiRNA agent has an asymmetric structure further containing 2 deoxyribonucleotides at the 3′ end of the antisense strand.

Exemplary α-1 antitrypsin targeting DsiRNA agents of the invention, and their associated α-1 antitrypsin target sequences, include the following, presented in the below series of tables:

Table Number:

(2) Selected Human Anti-α-1 antitrypsin DsiRNA Agents (Asymmetrics);

(3) Selected Human Anti-α-1 antitrypsin DsiRNAs, Unmodified Duplexes (Asymmetrics);

(4) DsiRNA Target Sequences (21mers) in α-1 antitrypsin mRNA;

(5) Selected Human Anti-α-1 antitrypsin “Blunt/Blunt” DsiRNAs; and

(6) DsiRNA Component 19 Nucleotide Target Sequences in α-1 antitrypsin mRNA

(7) Additional Human Anti-α-1 antitrypsin DsiRNA Agents (Asymmetrics);

(8) Additional Human Anti-α-1 antitrypsin DsiRNAs, Unmodified Duplexes (Asymmetrics);

(9) Additional DsiRNA Target Sequences (21mers) in α-1 antitrypsin mRNA;

(10) Additional Human Anti-α-1 antitrypsin “Blunt/Blunt” DsiRNAs; and

(11) Additional DsiRNA Component 19 Nucleotide Target Sequences in α-1 antitrypsin mRNA

(12) Further Human Anti-α-1 antitrypsin DsiRNA Agents (Asymmetrics);

(13) Further Human Anti-α-1 antitrypsin DsiRNAs, Unmodified Duplexes (Asymmetrics);

(14) Further DsiRNA Target Sequences (21mers) in α-1 antitrypsin mRNA;

(15) Further Human Anti-α-1 antitrypsin “Blunt/Blunt” DsiRNAs; and

(16) Further DsiRNA Component 19 Nucleotide Target Sequences in α-1 antitrypsin mRNA

(17) Other Human Anti-α-1 antitrypsin DsiRNA Agents (Asymmetrics);

(18) Other Human Anti-α-1 antitrypsin DsiRNAs, Unmodified Duplexes (Asymmetrics);

(19) Other DsiRNA Target Sequences (21mers) in α-1 antitrypsin mRNA;

(20) Other Human Anti-α-1 antitrypsin “Blunt/Blunt” DsiRNAs; and

(21) Other DsiRNA Component 19 Nucleotide Target Sequences in α-1 antitrypsin mRNA

TABLE 2 Selected Human Anti-α-1 antitrypsin DsiRNA Agents (Asymmetrics) 5′-CAUCCCACCAUGAUCAGGAUCACcc-3′(SEQ ID NO: 1)  3′-AUGUAGGGUGGUACUAGUCCUAGUGGG-5′(SEQ ID NO: 199)  AAT-395 Target: 5′-TACATCCCACCATGATCAGGATCACCC-3′(SEQ ID NO: 397)  5′-CAGCUGGCACACCAGUCCAACAGca-3′(SEQ ID NO: 2)  3′-CGGUCGACCGUGUGGUCAGGUUGUCGU-5′(SEQ ID NO: 200)  AAT-475 Target: 5′-GCCAGCTGGCACACCAGTCCAACAGCA-3′(SEQ ID NO: 398)  5′-GCUGGCACACCAGUCCAACAGCAcc-3′(SEQ ID NO: 3)  3′-GUCGACCGUGUGGUCAGGUUGUCGUGG-5′(SEQ ID NO: 201)  AAT-477 Target: 5′-CAGCTGGCACACCAGTCCAACAGCACC-3′(SEQ ID NO: 399)  5′-GGCACACCAGUCCAACAGCACCAat-3′(SEQ ID NO: 4)  3′-GACCGUGUGGUCAGGUUGUCGUGGUUA-5′(SEQ ID NO: 202)  AAT-480 Target: 5′-CTGGCACACCAGTCCAACAGCACCAAT-3′(SEQ ID NO: 400)  5′-GCACACCAGUCCAACAGCACCAAta-3′(SEQ ID NO: 5)  3′-ACCGUGUGGUCAGGUUGUCGUGGUUAU-5′(SEQ ID NO: 203)  AAT-481 Target: 5′-TGGCACACCAGTCCAACAGCACCAATA-3′(SEQ ID NO: 401)  5′-CACACCAGUCCAACAGCACCAAUat-3′(SEQ ID NO: 6)  3′-CCGUGUGGUCAGGUUGUCGUGGUUAUA-5′(SEQ ID NO: 204)  AAT-482 Target: 5′-GGCACACCAGTCCAACAGCACCAATAT-3′(SEQ ID NO: 402)  5′-ACACCAGUCCAACAGCACCAAUAtc-3′(SEQ ID NO: 7)  3′-CGUGUGGUCAGGUUGUCGUGGUUAUAG-5′(SEQ ID NO: 205)  AAT-483 Target: 5′-GCACACCAGTCCAACAGCACCAATATC-3′(SEQ ID NO: 403)  5′-CACCAGUCCAACAGCACCAAUAUct-3′(SEQ ID NO: 8)  3′-GUGUGGUCAGGUUGUCGUGGUUAUAGA-5′(SEQ ID NO: 206)  AAT-484 Target: 5′-CACACCAGTCCAACAGCACCAATATCT-3′(SEQ ID NO: 404)  5′-CCAAUAUCUUCUUCUCCCCAGUGag-3′(SEQ ID NO: 9)  3′-GUGGUUAUAGAAGAAGAGGGGUCACUC-5′(SEQ ID NO: 207)  AAT-500 Target: 5′-CACCAATATCTTCTTCTCCCCAGTGAG-3′(SEQ ID NO: 405)  5′-CAAUAUCUUCUUCUCCCCAGUGAgc-3′(SEQ ID NO: 10)  3′-UGGUUAUAGAAGAAGAGGGGUCACUCG-5′(SEQ ID NO: 208)  AAT-501 Target: 5′-ACCAATATCTTCTTCTCCCCAGTGAGC-3′(SEQ ID NO: 406)  5′-AAUAUCUUCUUCUCCCCAGUGAGca-3′(SEQ ID NO: 11)  3′-GGUUAUAGAAGAAGAGGGGUCACUCGU-5′(SEQ ID NO: 209)  AAT-502 Target: 5′-CCAATATCTTCTTCTCCCCAGTGAGCA-3′(SEQ ID NO: 407)  5′-AUAUCUUCUUCUCCCCAGUGAGCat-3′(SEQ ID NO: 12)  3′-GUUAUAGAAGAAGAGGGGUCACUCGUA-5′(SEQ ID NO: 210)  AAT-503 Target: 5′-CAATATCTTCTTCTCCCCAGTGAGCAT-3′(SEQ ID NO: 408)  5′-UAUCUUCUUCUCCCCAGUGAGCAtc-3′(SEQ ID NO: 13)  3′-UUAUAGAAGAAGAGGGGUCACUCGUAG-5′(SEQ ID NO: 211)  AAT-504 Target: 5′-AATATCTTCTTCTCCCCAGTGAGCATC-3′(SEQ ID NO: 409)  5′-AUCUUCUUCUCCCCAGUGAGCAUcg-3′(SEQ ID NO: 14)  3′-UAUAGAAGAAGAGGGGUCACUCGUAGC-5′(SEQ ID NO: 212)  AAT-505 Target: 5′-ATATCTTCTTCTCCCCAGTGAGCATCG-3′(SEQ ID NO: 410)  5′-UCUUCUUCUCCCCAGUGAGCAUCgc-3′(SEQ ID NO: 15)  3′-AUAGAAGAAGAGGGGUCACUCGUAGCG-5′(SEQ ID NO: 213)  AAT-506 Target: 5′-TATCTTCTTCTCCCCAGTGAGCATCGC-3′(SEQ ID NO: 411)  5′-CUUCUUCUCCCCAGUGAGCAUCGct-3′(SEQ ID NO: 16)  3′-UAGAAGAAGAGGGGUCACUCGUAGCGA-5′(SEQ ID NO: 214)  AAT-507 Target: 5′-ATCTTCTTCTCCCCAGTGAGCATCGCT-3′(SEQ ID NO: 412)  5′-UUCUUCUCCCCAGUGAGCAUCGCta-3′(SEQ ID NO: 17)  3′-AGAAGAAGAGGGGUCACUCGUAGCGAU-5′(SEQ ID NO: 215)  AAT-508 Target: 5′-TCTTCTTCTCCCCAGTGAGCATCGCTA-3′(SEQ ID NO: 413)  5′-UCUUCUCCCCAGUGAGCAUCGCUac-3′(SEQ ID NO: 18)  3′-GAAGAAGAGGGGUCACUCGUAGCGAUG-5′(SEQ ID NO: 216)  AAT-509 Target: 5′-CTTCTTCTCCCCAGTGAGCATCGCTAC-3′(SEQ ID NO: 414)  5′-CUUCUCCCCAGUGAGCAUCGCUAca-3′(SEQ ID NO: 19)  3′-AAGAAGAGGGGUCACUCGUAGCGAUGU-5′(SEQ ID NO: 217)  AAT-510 Target: 5′-TTCTTCTCCCCAGTGAGCATCGCTACA-3′(SEQ ID NO: 415)  5′-UCUCCCCAGUGAGCAUCGCUACAgc-3′(SEQ ID NO: 20)  3′-GAAGAGGGGUCACUCGUAGCGAUGUCG-5′(SEQ ID NO: 218)  AAT-512 Target: 5′-CTTCTCCCCAGTGAGCATCGCTACAGC-3′(SEQ ID NO: 416)  5′-CUCCCCAGUGAGCAUCGCUACAGcc-3′(SEQ ID NO: 21)  3′-AAGAGGGGUCACUCGUAGCGAUGUCGG-5′(SEQ ID NO: 219)  AAT-513 Target: 5′-TTCTCCCCAGTGAGCATCGCTACAGCC-3′(SEQ ID NO: 417)  5′-CCCCAGUGAGCAUCGCUACAGCCtt-3′(SEQ ID NO: 22)  3′-GAGGGGUCACUCGUAGCGAUGUCGGAA-5′(SEQ ID NO: 220)  AAT-515 Target: 5′-CTCCCCAGTGAGCATCGCTACAGCCTT-3′(SEQ ID NO: 418)  5′-ACAGCCUUUGCAAUGCUCUCCCUgg-3′(SEQ ID NO: 23)  3′-GAUGUCGGAAACGUUACGAGAGGGACC-5′(SEQ ID NO: 221)  AAT-532 Target: 5′-CTACAGCCTTTGCAATGCTCTCCCTGG-3′(SEQ ID NO: 419)  5′-UGCAAUGCUCUCCCUGGGGACCAag-3′(SEQ ID NO: 24)  3′-AAACGUUACGAGAGGGACCCCUGGUUC-5′(SEQ ID NO: 222)  AAT-540 Target: 5′-TTTGCAATGCTCTCCCTGGGGACCAAG-3′(SEQ ID NO: 420)  5′-AAAUCCUGGAGGGCCUGAAUUUCaa-3′(SEQ ID NO: 25)  3′-ACUUUAGGACCUCCCGGACUUAAAGUU-5′(SEQ ID NO: 223)  AAT-581 Target: 5′-TGAAATCCTGGAGGGCCTGAATTTCAA-3′(SEQ ID NO: 421)  5′-AAUCCUGGAGGGCCUGAAUUUCAac-3′(SEQ ID NO: 26)  3′-CUUUAGGACCUCCCGGACUUAAAGUUG-5′(SEQ ID NO: 224)  AAT-582 Target: 5′-GAAATCCTGGAGGGCCTGAATTTCAAC-3′(SEQ ID NO: 422)  5′-AUCCUGGAGGGCCUGAAUUUCAAcc-3′(SEQ ID NO: 27)  3′-UUUAGGACCUCCCGGACUUAAAGUUGG-5′(SEQ ID NO: 225)  AAT-583 Target: 5′-AAATCCTGGAGGGCCTGAATTTCAACC-3′(SEQ ID NO: 423)  5′-CCUGGAGGGCCUGAAUUUCAACCtc-3′(SEQ ID NO: 28)  3′-UAGGACCUCCCGGACUUAAAGUUGGAG-5′(SEQ ID NO: 226)  AAT-585 Target: 5′-ATCCTGGAGGGCCTGAATTTCAACCTC-3′(SEQ ID NO: 424)  5′-CUGGAGGGCCUGAAUUUCAACCUca-3′(SEQ ID NO: 29)  3′-AGGACCUCCCGGACUUAAAGUUGGAGU-5′(SEQ ID NO: 227)  AAT-586 Target: 5′-TCCTGGAGGGCCTGAATTTCAACCTCA-3′(SEQ ID NO: 425)  5′-UGGAGGGCCUGAAUUUCAACCUCac-3′(SEQ ID NO: 30)  3′-GGACCUCCCGGACUUAAAGUUGGAGUG-5′(SEQ ID NO: 228)  AAT-587 Target: 5′-CCTGGAGGGCCTGAATTTCAACCTCAC-3′(SEQ ID NO: 426)  5′-CAUGAAGGCUUCCAGGAACUCCUcc-3′(SEQ ID NO: 31)  3′-AGGUACUUCCGAAGGUCCUUGAGGAGG-5′(SEQ ID NO: 229)  AAT-634 Target: 5′-TCCATGAAGGCTTCCAGGAACTCCTCC-3′(SEQ ID NO: 427)  5′-GAAGGCUUCCAGGAACUCCUCCGta-3′(SEQ ID NO: 32)  3′-UACUUCCGAAGGUCCUUGAGGAGGCAU-5′(SEQ ID NO: 230)  AAT-637 Target: 5′-ATGAAGGCTTCCAGGAACTCCTCCGTA-3′(SEQ ID NO: 428)  5′-AAGGCUUCCAGGAACUCCUCCGUac-3′(SEQ ID NO: 33)  3′-ACUUCCGAAGGUCCUUGAGGAGGCAUG-5′(SEQ ID NO: 231)  AAT-638 Target: 5′-TGAAGGCTTCCAGGAACTCCTCCGTAC-3′(SEQ ID NO: 429)  5′-AGCCAGACAGCCAGCUCCAGCUGac-3′(SEQ ID NO: 34)  3′-GGUCGGUCUGUCGGUCGAGGUCGACUG-5′(SEQ ID NO: 232)  AAT-671 Target: 5′-CCAGCCAGACAGCCAGCTCCAGCTGAC-3′(SEQ ID NO: 430)  5′-CCAGACAGCCAGCUCCAGCUGACca-3′(SEQ ID NO: 35)  3′-UCGGUCUGUCGGUCGAGGUCGACUGGU-5′(SEQ ID NO: 233)  AAT-673 Target: 5′-AGCCAGACAGCCAGCTCCAGCTGACCA-3′(SEQ ID NO: 431)  5′-AGACAGCCAGCUCCAGCUGACCAcc-3′(SEQ ID NO: 36)  3′-GGUCUGUCGGUCGAGGUCGACUGGUGG-5′(SEQ ID NO: 234)  AAT-675 Target: 5′-CCAGACAGCCAGCTCCAGCTGACCACC-3′(SEQ ID NO: 432)  5′-GACAGCCAGCUCCAGCUGACCACcg-3′(SEQ ID NO: 37)  3′-GUCUGUCGGUCGAGGUCGACUGGUGGC-5′(SEQ ID NO: 235)  AAT-676 Target: 5′-CAGACAGCCAGCTCCAGCTGACCACCG-3′(SEQ ID NO: 433)  5′-UAGUGGAUAAGUUUUUGGAGGAUgt-3′(SEQ ID NO: 38)  3′-CGAUCACCUAUUCAAAAACCUCCUACA-5′(SEQ ID NO: 236)  AAT-734 Target: 5′-GCTAGTGGATAAGTTTTTGGAGGATGT-3′(SEQ ID NO: 434)  5′-AGUGGAUAAGUUUUUGGAGGAUGtt-3′(SEQ ID NO: 39)  3′-GAUCACCUAUUCAAAAACCUCCUACAA-5′(SEQ ID NO: 237)  AAT-735 Target: 5′-CTAGTGGATAAGTTTTTGGAGGATGTT-3′(SEQ ID NO: 435)  5′-GUGGAUAAGUUUUUGGAGGAUGUta-3′(SEQ ID NO: 40)  3′-AUCACCUAUUCAAAAACCUCCUACAAU-5′(SEQ ID NO: 238)  AAT-736 Target: 5′-TAGTGGATAAGTTTTTGGAGGATGTTA-3′(SEQ ID NO: 436)  5′-UGGAUAAGUUUUUGGAGGAUGUUaa-3′(SEQ ID NO: 41)  3′-UCACCUAUUCAAAAACCUCCUACAAUU-5′(SEQ ID NO: 239)  AAT-737 Target: 5′-AGTGGATAAGTTTTTGGAGGATGTTAA-3′(SEQ ID NO: 437)  5′-GGAUAAGUUUUUGGAGGAUGUUAaa-3′(SEQ ID NO: 42)  3′-CACCUAUUCAAAAACCUCCUACAAUUU-5′(SEQ ID NO: 240)  AAT-738 Target: 5′-GTGGATAAGTTTTTGGAGGATGTTAAA-3′(SEQ ID NO: 438)  5′-GAUAAGUUUUUGGAGGAUGUUAAaa-3′(SEQ ID NO: 43)  3′-ACCUAUUCAAAAACCUCCUACAAUUUU-5′(SEQ ID NO: 241)  AAT-739 Target: 5′-TGGATAAGTTTTTGGAGGATGTTAAAA-3′(SEQ ID NO: 439)  5′-AUAAGUUUUUGGAGGAUGUUAAAaa-3′(SEQ ID NO: 44)  3′-CCUAUUCAAAAACCUCCUACAAUUUUU-5′(SEQ ID NO: 242)  AAT-740 Target: 5′-GGATAAGTTTTTGGAGGATGTTAAAAA-3′(SEQ ID NO: 440)  5′-UGUACCACUCAGAAGCCUUCACUgt-3′(SEQ ID NO: 45)  3′-CAACAUGGUGAGUCUUCGGAAGUGACA-5′(SEQ ID NO: 243)  AAT-767 Target: 5′-GTTGTACCACTCAGAAGCCTTCACTGT-3′(SEQ ID NO: 441)  5′-GUACCACUCAGAAGCCUUCACUGtc-3′(SEQ ID NO: 46)  3′-AACAUGGUGAGUCUUCGGAAGUGACAG-5′(SEQ ID NO: 244)  AAT-768 Target: 5′-TTGTACCACTCAGAAGCCTTCACTGTC-3′(SEQ ID NO: 442)  5′-ACUCAAGGGAAAAUUGUGGAUUUgg-3′(SEQ ID NO: 47)  3′-CAUGAGUUCCCUUUUAACACCUAAACC-5′(SEQ ID NO: 245)  AAT-850 Target: 5′-GTACTCAAGGGAAAATTGTGGATTTGG-3′(SEQ ID NO: 443)  5′-CUCAAGGGAAAAUUGUGGAUUUGgt-3′(SEQ ID NO: 48)  3′-AUGAGUUCCCUUUUAACACCUAAACCA-5′(SEQ ID NO: 246)  AAT-851 Target: 5′-TACTCAAGGGAAAATTGTGGATTTGGT-3′(SEQ ID NO: 444)  5′-UCAAGGGAAAAUUGUGGAUUUGGtc-3′(SEQ ID NO: 49)  3′-UGAGUUCCCUUUUAACACCUAAACCAG-5′(SEQ ID NO: 247)  AAT-852 Target: 5′-ACTCAAGGGAAAATTGTGGATTTGGTC-3′(SEQ ID NO: 445)  5′-CAAGGGAAAAUUGUGGAUUUGGUca-3′(SEQ ID NO: 50)  3′-GAGUUCCCUUUUAACACCUAAACCAGU-5′(SEQ ID NO: 248)  AAT-853 Target: 5′-CTCAAGGGAAAATTGTGGATTTGGTCA-3′(SEQ ID NO: 446)  5′-AAGGGAAAAUUGUGGAUUUGGUCaa-3′(SEQ ID NO: 51)  3′-AGUUCCCUUUUAACACCUAAACCAGUU-5′(SEQ ID NO: 249)  AAT-854 Target: 5′-TCAAGGGAAAATTGTGGATTTGGTCAA-3′(SEQ ID NO: 447)  5′-AGGGAAAAUUGUGGAUUUGGUCAag-3′(SEQ ID NO: 52)  3′-GUUCCCUUUUAACACCUAAACCAGUUC-5′(SEQ ID NO: 250)  AAT-855 Target: 5′-CAAGGGAAAATTGTGGATTTGGTCAAG-3′(SEQ ID NO: 448)  5′-GGGAAAAUUGUGGAUUUGGUCAAgg-3′(SEQ ID NO: 53)  3′-UUCCCUUUUAACACCUAAACCAGUUCC-5′(SEQ ID NO: 251)  AAT-856 Target: 5′-AAGGGAAAATTGTGGATTTGGTCAAGG-3′(SEQ ID NO: 449)  5′-GGAAAAUUGUGGAUUUGGUCAAGga-3′(SEQ ID NO: 54)  3′-UCCCUUUUAACACCUAAACCAGUUCCU-5′(SEQ ID NO: 252)  AAT-857 Target: 5′-AGGGAAAATTGTGGATTTGGTCAAGGA-3′(SEQ ID NO: 450)  5′-GAAAAUUGUGGAUUUGGUCAAGGag-3′(SEQ ID NO: 55)  3′-CCCUUUUAACACCUAAACCAGUUCCUC-5′(SEQ ID NO: 253)  AAT-858 Target: 5′-GGGAAAATTGTGGATTTGGTCAAGGAG-3′(SEQ ID NO: 451)  5′-AAAAUUGUGGAUUUGGUCAAGGAgc-3′(SEQ ID NO: 56)  3′-CCUUUUAACACCUAAACCAGUUCCUCG-5′(SEQ ID NO: 254)  AAT-859 Target: 5′-GGAAAATTGTGGATTTGGTCAAGGAGC-3′(SEQ ID NO: 452)  5′-AAAUUGUGGAUUUGGUCAAGGAGct-3′(SEQ ID NO: 57)  3′-CUUUUAACACCUAAACCAGUUCCUCGA-5′(SEQ ID NO: 255)  AAT-860 Target: 5′-GAAAATTGTGGATTTGGTCAAGGAGCT-3′(SEQ ID NO: 453)  5′-AAUUGUGGAUUUGGUCAAGGAGCtt-3′(SEQ ID NO: 58)  3′-UUUUAACACCUAAACCAGUUCCUCGAA-5′(SEQ ID NO: 256)  AAT-861 Target: 5′-AAAATTGTGGATTTGGTCAAGGAGCTT-3′(SEQ ID NO: 454)  5′-AUUGUGGAUUUGGUCAAGGAGCUtg-3′(SEQ ID NO: 59)  3′-UUUAACACCUAAACCAGUUCCUCGAAC-5′(SEQ ID NO: 257)  AAT-862 Target: 5′-AAATTGTGGATTTGGTCAAGGAGCTTG-3′(SEQ ID NO: 455)  5′-UUGUGGAUUUGGUCAAGGAGCUUga-3′(SEQ ID NO: 60)  3′-UUAACACCUAAACCAGUUCCUCGAACU-5′(SEQ ID NO: 258)  AAT-863 Target: 5′-AATTGTGGATTTGGTCAAGGAGCTTGA-3′(SEQ ID NO: 456)  5′-UGUGGAUUUGGUCAAGGAGCUUGac-3′(SEQ ID NO: 61)  3′-UAACACCUAAACCAGUUCCUCGAACUG-5′(SEQ ID NO: 259)  AAT-864 Target: 5′-ATTGTGGATTTGGTCAAGGAGCTTGAC-3′(SEQ ID NO: 457)  5′-GUGGAUUUGGUCAAGGAGCUUGAca-3′(SEQ ID NO: 62)  3′-AACACCUAAACCAGUUCCUCGAACUGU-5′(SEQ ID NO: 260)  AAT-865 Target: 5′-TTGTGGATTTGGTCAAGGAGCTTGACA-3′(SEQ ID NO: 458)  5′-UGGAUUUGGUCAAGGAGCUUGACag-3′(SEQ ID NO: 63)  3′-ACACCUAAACCAGUUCCUCGAACUGUC-5′(SEQ ID NO: 261)  AAT-866 Target: 5′-TGTGGATTTGGTCAAGGAGCTTGACAG-3′(SEQ ID NO: 459)  5′-GGAUUUGGUCAAGGAGCUUGACAga-3′(SEQ ID NO: 64)  3′-CACCUAAACCAGUUCCUCGAACUGUCU-5′(SEQ ID NO: 262)  AAT-867 Target: 5′-GTGGATTTGGTCAAGGAGCTTGACAGA-3′(SEQ ID NO: 460)  5′-GAUUUGGUCAAGGAGCUUGACAGag-3′(SEQ ID NO: 65)  3′-ACCUAAACCAGUUCCUCGAACUGUCUC-5′(SEQ ID NO: 263)  AAT-868 Target: 5′-TGGATTTGGTCAAGGAGCTTGACAGAG-3′(SEQ ID NO: 461)  5′-AUUUGGUCAAGGAGCUUGACAGAga-3′(SEQ ID NO: 66)  3′-CCUAAACCAGUUCCUCGAACUGUCUCU-5′(SEQ ID NO: 264)  AAT-869 Target: 5′-GGATTTGGTCAAGGAGCTTGACAGAGA-3′(SEQ ID NO: 462)  5′-UUUGGUCAAGGAGCUUGACAGAGac-3′(SEQ ID NO: 67)  3′-CUAAACCAGUUCCUCGAACUGUCUCUG-5′(SEQ ID NO: 265)  AAT-870 Target: 5′-GATTTGGTCAAGGAGCTTGACAGAGAC-3′(SEQ ID NO: 463)  5′-UUGGUCAAGGAGCUUGACAGAGAca-3′(SEQ ID NO: 68)  3′-UAAACCAGUUCCUCGAACUGUCUCUGU-5′(SEQ ID NO: 266)  AAT-871 Target: 5′-ATTTGGTCAAGGAGCTTGACAGAGACA-3′(SEQ ID NO: 464)  5′-UGGUCAAGGAGCUUGACAGAGACac-3′(SEQ ID NO: 69)  3′-AAACCAGUUCCUCGAACUGUCUCUGUG-5′(SEQ ID NO: 267)  AAT-872 Target: 5′-TTTGGTCAAGGAGCTTGACAGAGACAC-3′(SEQ ID NO: 465)  5′-CAGUUUUUGCUCUGGUGAAUUACat-3′(SEQ ID NO: 70)  3′-GUGUCAAAAACGAGACCACUUAAUGUA-5′(SEQ ID NO: 268)  AAT-896 Target: 5′-CACAGTTTTTGCTCTGGTGAATTACAT-3′(SEQ ID NO: 466)  5′-AGUUUUUGCUCUGGUGAAUUACAtc-3′(SEQ ID NO: 71)  3′-UGUCAAAAACGAGACCACUUAAUGUAG-5′(SEQ ID NO: 269)  AAT-897 Target: 5′-ACAGTTTTTGCTCTGGTGAATTACATC-3′(SEQ ID NO: 467)  5′-GUUUUUGCUCUGGUGAAUUACAUct-3′(SEQ ID NO: 72)  3′-GUCAAAAACGAGACCACUUAAUGUAGA-5′(SEQ ID NO: 270)  AAT-898 Target: 5′-CAGTTTTTGCTCTGGTGAATTACATCT-3′(SEQ ID NO: 468)  5′-UUUUUGCUCUGGUGAAUUACAUCtt-3′(SEQ ID NO: 73)  3′-UCAAAAACGAGACCACUUAAUGUAGAA-5′(SEQ ID NO: 271)  AAT-899 Target: 5′-AGTTTTTGCTCTGGTGAATTACATCTT-3′(SEQ ID NO: 469)  5′-AAAGGCAAAUGGGAGAGACCCUUtg-3′(SEQ ID NO: 74)  3′-AAUUUCCGUUUACCCUCUCUGGGAAAC-5′(SEQ ID NO: 272)  AAT-928 Target: 5′-TTAAAGGCAAATGGGAGAGACCCTTTG-3′(SEQ ID NO: 470)  5′-AAGGCAAAUGGGAGAGACCCUUUga-3′(SEQ ID NO: 75)  3′-AUUUCCGUUUACCCUCUCUGGGAAACU-5′(SEQ ID NO: 273)  AAT-929 Target: 5′-TAAAGGCAAATGGGAGAGACCCTTTGA-3′(SEQ ID NO: 471)  5′-AGGCAAAUGGGAGAGACCCUUUGaa-3′(SEQ ID NO: 76)  3′-UUUCCGUUUACCCUCUCUGGGAAACUU-5′(SEQ ID NO: 274)  AAT-930 Target: 5′-AAAGGCAAATGGGAGAGACCCTTTGAA-3′(SEQ ID NO: 472)  5′-GGCAAAUGGGAGAGACCCUUUGAag-3′(SEQ ID NO: 77)  3′-UUCCGUUUACCCUCUCUGGGAAACUUC-5′(SEQ ID NO: 275)  AAT-931 Target: 5′-AAGGCAAATGGGAGAGACCCTTTGAAG-3′(SEQ ID NO: 473)  5′-AGGAAGAGGACUUCCACGUGGACca-3′(SEQ ID NO: 78)  3′-GCUCCUUCUCCUGAAGGUGCACCUGGU-5′(SEQ ID NO: 276)  AAT-968 Target: 5′-CGAGGAAGAGGACTTCCACGTGGACCA-3′(SEQ ID NO: 474)  5′-GGAAGAGGACUUCCACGUGGACCag-3′(SEQ ID NO: 79)  3′-CUCCUUCUCCUGAAGGUGCACCUGGUC-5′(SEQ ID NO: 277)  AAT-969 Target: 5′-GAGGAAGAGGACTTCCACGTGGACCAG-3′(SEQ ID NO: 475)  5′-GAAGAGGACUUCCACGUGGACCAgg-3′(SEQ ID NO: 80)  3′-UCCUUCUCCUGAAGGUGCACCUGGUCC-5′(SEQ ID NO: 278)  AAT-970 Target: 5′-AGGAAGAGGACTTCCACGTGGACCAGG-3′(SEQ ID NO: 476)  5′-AAGAGGACUUCCACGUGGACCAGgt-3′(SEQ ID NO: 81)  3′-CCUUCUCCUGAAGGUGCACCUGGUCCA-5′(SEQ ID NO: 279)  AAT-971 Target: 5′-GGAAGAGGACTTCCACGTGGACCAGGT-3′(SEQ ID NO: 477)  5′-GAGGACUUCCACGUGGACCAGGUga-3′(SEQ ID NO: 82)  3′-UUCUCCUGAAGGUGCACCUGGUCCACU-5′(SEQ ID NO: 280)  AAT-973 Target: 5′-AAGAGGACTTCCACGTGGACCAGGTGA-3′(SEQ ID NO: 478)  5′-AGGACUUCCACGUGGACCAGGUGac-3′(SEQ ID NO: 83)  3′-UCUCCUGAAGGUGCACCUGGUCCACUG-5′(SEQ ID NO: 281)  AAT-974 Target: 5′-AGAGGACTTCCACGTGGACCAGGTGAC-3′(SEQ ID NO: 479)  5′-GACUUCCACGUGGACCAGGUGACca-3′(SEQ ID NO: 84)  3′-UCCUGAAGGUGCACCUGGUCCACUGGU-5′(SEQ ID NO: 282)  AAT-976 Target: 5′-AGGACTTCCACGTGGACCAGGTGACCA-3′(SEQ ID NO: 480)  5′-GUUUAGGCAUGUUUAACAUCCAGca-3′(SEQ ID NO: 85)  3′-CGCAAAUCCGUACAAAUUGUAGGUCGU-5′(SEQ ID NO: 283)  AAT-1025 Target: 5′-GCGTTTAGGCATGTTTAACATCCAGCA-3′(SEQ ID NO: 481)  5′-UUUAGGCAUGUUUAACAUCCAGCac-3′(SEQ ID NO: 86)  3′-GCAAAUCCGUACAAAUUGUAGGUCGUG-5′(SEQ ID NO: 284)  AAT-1026 Target: 5′-CGTTTAGGCATGTTTAACATCCAGCAC-3′(SEQ ID NO: 482)  5′-GCUGUCCAGCUGGGUGCUGCUGAtg-3′(SEQ ID NO: 87)  3′-UUCGACAGGUCGACCCACGACGACUAC-5′(SEQ ID NO: 285)  AAT-1059 Target: 5′-AAGCTGTCCAGCTGGGTGCTGCTGATG-3′(SEQ ID NO: 483)  5′-CUGUCCAGCUGGGUGCUGCUGAUga-3′(SEQ ID NO: 88)  3′-UCGACAGGUCGACCCACGACGACUACU-5′(SEQ ID NO: 286)  AAT-1060 Target: 5′-AGCTGTCCAGCTGGGTGCTGCTGATGA-3′(SEQ ID NO: 484)  5′-CAAUGCCACCGCCAUCUUCUUCCtg-3′(SEQ ID NO: 89)  3′-CCGUUACGGUGGCGGUAGAAGAAGGAC-5′(SEQ ID NO: 287)  AAT-1095 Target: 5′-GGCAATGCCACCGCCATCTTCTTCCTG-3′(SEQ ID NO: 485)  5′-AAUGCCACCGCCAUCUUCUUCCUgc-3′(SEQ ID NO: 90)  3′-CGUUACGGUGGCGGUAGAAGAAGGACG-5′(SEQ ID NO: 288)  AAT-1096 Target: 5′-GCAATGCCACCGCCATCTTCTTCCTGC-3′(SEQ ID NO: 486)  5′-CCACCGCCAUCUUCUUCCUGCCUga-3′(SEQ ID NO: 91)  3′-ACGGUGGCGGUAGAAGAAGGACGGACU-5′(SEQ ID NO: 289)  AAT-1100 Target: 5′-TGCCACCGCCATCTTCTTCCTGCCTGA-3′(SEQ ID NO: 487)  5′-CACCGCCAUCUUCUUCCUGCCUGat-3′(SEQ ID NO: 92)  3′-CGGUGGCGGUAGAAGAAGGACGGACUA-5′(SEQ ID NO: 290)  AAT-1101 Target: 5′-GCCACCGCCATCTTCTTCCTGCCTGAT-3′(SEQ ID NO: 488)  5′-ACCGCCAUCUUCUUCCUGCCUGAtg-3′(SEQ ID NO: 93)  3′-GGUGGCGGUAGAAGAAGGACGGACUAC-5′(SEQ ID NO: 291)  AAT-1102 Target: 5′-CCACCGCCATCTTCTTCCTGCCTGATG-3′(SEQ ID NO: 489)  5′-CCGCCAUCUUCUUCCUGCCUGAUga-3′(SEQ ID NO: 94)  3′-GUGGCGGUAGAAGAAGGACGGACUACU-5′(SEQ ID NO: 292)  AAT-1103 Target: 5′-CACCGCCATCTTCTTCCTGCCTGATGA-3′(SEQ ID NO: 490)  5′-CGCCAUCUUCUUCCUGCCUGAUGag-3′(SEQ ID NO: 95)  3′-UGGCGGUAGAAGAAGGACGGACUACUC-5′(SEQ ID NO: 293)  AAT-1104 Target: 5′-ACCGCCATCTTCTTCCTGCCTGATGAG-3′(SEQ ID NO: 491)  5′-GCCAUCUUCUUCCUGCCUGAUGAgg-3′(SEQ ID NO: 96)  3′-GGCGGUAGAAGAAGGACGGACUACUCC-5′(SEQ ID NO: 294)  AAT-1105 Target: 5′-CCGCCATCTTCTTCCTGCCTGATGAGG-3′(SEQ ID NO: 492)  5′-AUCUUCUUCCUGCCUGAUGAGGGga-3′(SEQ ID NO: 97)  3′-GGUAGAAGAAGGACGGACUACUCCCCU-5′(SEQ ID NO: 295)  AAT-1108 Target: 5′-CCATCTTCTTCCTGCCTGATGAGGGGA-3′(SEQ ID NO: 493)  5′-CUUCCUGCCUGAUGAGGGGAAACta-3′(SEQ ID NO: 98)  3′-AAGAAGGACGGACUACUCCCCUUUGAU-5′(SEQ ID NO: 296)  AAT-1113 Target: 5′-TTCTTCCTGCCTGATGAGGGGAAACTA-3′(SEQ ID NO: 494)  5′-UUCCUGCCUGAUGAGGGGAAACUac-3′(SEQ ID NO: 99)  3′-AGAAGGACGGACUACUCCCCUUUGAUG-5′(SEQ ID NO: 297)  AAT-1114 Target: 5′-TCTTCCTGCCTGATGAGGGGAAACTAC-3′(SEQ ID NO: 495)  5′-UCCUGCCUGAUGAGGGGAAACUAca-3′(SEQ ID NO: 100)  3′-GAAGGACGGACUACUCCCCUUUGAUGU-5′(SEQ ID NO: 298)  AAT-1115 Target: 5′-CTTCCTGCCTGATGAGGGGAAACTACA-3′(SEQ ID NO: 496)  5′-CCUGCCUGAUGAGGGGAAACUACag-3′(SEQ ID NO: 101)  3′-AAGGACGGACUACUCCCCUUUGAUGUC-5′(SEQ ID NO: 299)  AAT-1116 Target: 5′-TTCCTGCCTGATGAGGGGAAACTACAG-3′(SEQ ID NO: 497)  5′-CUGCCUGAUGAGGGGAAACUACAgc-3′(SEQ ID NO: 102)  3′-AGGACGGACUACUCCCCUUUGAUGUCG-5′(SEQ ID NO: 300)  AAT-1117 Target: 5′-TCCTGCCTGATGAGGGGAAACTACAGC-3′(SEQ ID NO: 498)  5′-UGCCUGAUGAGGGGAAACUACAGca-3′(SEQ ID NO: 103)  3′-GGACGGACUACUCCCCUUUGAUGUCGU-5′(SEQ ID NO: 301)  AAT-1118 Target: 5′-CCTGCCTGATGAGGGGAAACTACAGCA-3′(SEQ ID NO: 499)  5′-AGCACCUGGAAAAUGAACUCACCca-3′(SEQ ID NO: 104)  3′-UGUCGUGGACCUUUUACUUGAGUGGGU-5′(SEQ ID NO: 302)  AAT-1139 Target: 5′-ACAGCACCTGGAAAATGAACTCACCCA-3′(SEQ ID NO: 500)  5′-GCACCUGGAAAAUGAACUCACCCac-3′(SEQ ID NO: 105)  3′-GUCGUGGACCUUUUACUUGAGUGGGUG-5′(SEQ ID NO: 303)  AAT-1140 Target: 5′-CAGCACCTGGAAAATGAACTCACCCAC-3′(SEQ ID NO: 501)  5′-CACCUGGAAAAUGAACUCACCCAcg-3′(SEQ ID NO: 106)  3′-UCGUGGACCUUUUACUUGAGUGGGUGC-5′(SEQ ID NO: 304)  AAT-1141 Target: 5′-AGCACCTGGAAAATGAACTCACCCACG-3′(SEQ ID NO: 502)  5′-ACCUGGAAAAUGAACUCACCCACga-3′(SEQ ID NO: 107)  3′-CGUGGACCUUUUACUUGAGUGGGUGCU-5′(SEQ ID NO: 305)  AAT-1142 Target: 5′-GCACCTGGAAAATGAACTCACCCACGA-3′(SEQ ID NO: 503)  5′-CCUGGAAAAUGAACUCACCCACGat-3′(SEQ ID NO: 108)  3′-GUGGACCUUUUACUUGAGUGGGUGCUA-5′(SEQ ID NO: 306)  AAT-1143 Target: 5′-CACCTGGAAAATGAACTCACCCACGAT-3′(SEQ ID NO: 504)  5′-AUAUCAUCACCAAGUUCCUGGAAaa-3′(SEQ ID NO: 109)  3′-GCUAUAGUAGUGGUUCAAGGACCUUUU-5′(SEQ ID NO: 307)  AAT-1166 Target: 5′-CGATATCATCACCAAGTTCCTGGAAAA-3′(SEQ ID NO: 505)  5′-UAUCAUCACCAAGUUCCUGGAAAat-3′(SEQ ID NO: 110)  3′-CUAUAGUAGUGGUUCAAGGACCUUUUA-5′(SEQ ID NO: 308)  AAT-1167 Target: 5′-GATATCATCACCAAGTTCCTGGAAAAT-3′(SEQ ID NO: 506)  5′-AUCAUCACCAAGUUCCUGGAAAAtg-3′(SEQ ID NO: 111)  3′-UAUAGUAGUGGUUCAAGGACCUUUUAC-5′(SEQ ID NO: 309)  AAT-1168 Target: 5′-ATATCATCACCAAGTTCCTGGAAAATG-3′(SEQ ID NO: 507)  5′-UCAUCACCAAGUUCCUGGAAAAUga-3′(SEQ ID NO: 112)  3′-AUAGUAGUGGUUCAAGGACCUUUUACU-5′(SEQ ID NO: 310)  AAT-1169 Target: 5′-TATCATCACCAAGTTCCTGGAAAATGA-3′(SEQ ID NO: 508)  5′-CAUCACCAAGUUCCUGGAAAAUGaa-3′(SEQ ID NO: 113)  3′-UAGUAGUGGUUCAAGGACCUUUUACUU-5′(SEQ ID NO: 311)  AAT-1170 Target: 5′-ATCATCACCAAGTTCCTGGAAAATGAA-3′(SEQ ID NO: 509)  5′-AUCACCAAGUUCCUGGAAAAUGAag-3′(SEQ ID NO: 114)  3′-AGUAGUGGUUCAAGGACCUUUUACUUC-5′(SEQ ID NO: 312)  AAT-1171 Target: 5′-TCATCACCAAGTTCCTGGAAAATGAAG-3′(SEQ ID NO: 510)  5′-UCACCAAGUUCCUGGAAAAUGAAga-3′(SEQ ID NO: 115)  3′-GUAGUGGUUCAAGGACCUUUUACUUCU-5′(SEQ ID NO: 313)  AAT-1172 Target: 5′-CATCACCAAGTTCCTGGAAAATGAAGA-3′(SEQ ID NO: 511)  5′-CACCAAGUUCCUGGAAAAUGAAGac-3′(SEQ ID NO: 116)  3′-UAGUGGUUCAAGGACCUUUUACUUCUG-5′(SEQ ID NO: 314)  AAT-1173 Target: 5′-ATCACCAAGTTCCTGGAAAATGAAGAC-3′(SEQ ID NO: 512)  5′-ACCAAGUUCCUGGAAAAUGAAGAca-3′(SEQ ID NO: 117)  3′-AGUGGUUCAAGGACCUUUUACUUCUGU-5′(SEQ ID NO: 315)  AAT-1174 Target: 5′-TCACCAAGTTCCTGGAAAATGAAGACA-3′(SEQ ID NO: 513)  5′-CCAAGUUCCUGGAAAAUGAAGACag-3′(SEQ ID NO: 118)  3′-GUGGUUCAAGGACCUUUUACUUCUGUC-5′(SEQ ID NO: 316)  AAT-1175 Target: 5′-CACCAAGTTCCTGGAAAATGAAGACAG-3′(SEQ ID NO: 514)  5′-AGGUCUUCAGCAAUGGGGCUGACct-3′(SEQ ID NO: 119)  3′-AUUCCAGAAGUCGUUACCCCGACUGGA-5′(SEQ ID NO: 317)  AAT-1286 Target: 5′-TAAGGTCTTCAGCAATGGGGCTGACCT-3′(SEQ ID NO: 515)  5′-CAAUGGGGCUGACCUCUCCGGGGtc-3′(SEQ ID NO: 120)  3′-UCGUUACCCCGACUGGAGAGGCCCCAG-5′(SEQ ID NO: 318)  AAT-1296 Target: 5′-AGCAATGGGGCTGACCTCTCCGGGGTC-3′(SEQ ID NO: 516)  5′-AAUGGGGCUGACCUCUCCGGGGUca-3′(SEQ ID NO: 121)  3′-CGUUACCCCGACUGGAGAGGCCCCAGU-5′(SEQ ID NO: 319)  AAT-1297 Target: 5′-GCAATGGGGCTGACCTCTCCGGGGTCA-3′(SEQ ID NO: 517)  5′-AUGGGGCUGACCUCUCCGGGGUCac-3′(SEQ ID NO: 122)  3′-GUUACCCCGACUGGAGAGGCCCCAGUG-5′(SEQ ID NO: 320)  AAT-1298 Target: 5′-CAATGGGGCTGACCTCTCCGGGGTCAC-3′(SEQ ID NO: 518)  5′-GAGGAGGCACCCCUGAAGCUCUCca-3′(SEQ ID NO: 123)  3′-GUCUCCUCCGUGGGGACUUCGAGAGGU-5′(SEQ ID NO: 321)  AAT-1324 Target: 5′-CAGAGGAGGCACCCCTGAAGCTCTCCA-3′(SEQ ID NO: 519)  5′-GGAGGCACCCCUGAAGCUCUCCAag-3′(SEQ ID NO: 124)  3′-CUCCUCCGUGGGGACUUCGAGAGGUUC-5′(SEQ ID NO: 322)  AAT-1326 Target: 5′-GAGGAGGCACCCCTGAAGCTCTCCAAG-3′(SEQ ID NO: 520)  5′-CUGAAGCUCUCCAAGGCCGUGCAta-3′(SEQ ID NO: 125)  3′-GGGACUUCGAGAGGUUCCGGCACGUAU-5′(SEQ ID NO: 323)  AAT-1336 Target: 5′-CCCTGAAGCTCTCCAAGGCCGTGCATA-3′(SEQ ID NO: 521)  5′-CGUGCAUAAGGCUGUGCUGACCAtc-3′(SEQ ID NO: 126)  3′-CGGCACGUAUUCCGACACGACUGGUAG-5′(SEQ ID NO: 324)  AAT-1353 Target: 5′-GCCGTGCATAAGGCTGTGCTGACCATC-3′(SEQ ID NO: 522)  5′-GUGCAUAAGGCUGUGCUGACCAUcg-3′(SEQ ID NO: 127)  3′-GGCACGUAUUCCGACACGACUGGUAGC-5′(SEQ ID NO: 325)  AAT-1354 Target: 5′-CCGTGCATAAGGCTGTGCTGACCATCG-3′(SEQ ID NO: 523)  5′-UGCAUAAGGCUGUGCUGACCAUCga-3′(SEQ ID NO: 128)  3′-GCACGUAUUCCGACACGACUGGUAGCU-5′(SEQ ID NO: 326)  AAT-1355 Target: 5′-CGTGCATAAGGCTGTGCTGACCATCGA-3′(SEQ ID NO: 524)  5′-GCAUAAGGCUGUGCUGACCAUCGac-3′(SEQ ID NO: 129)  3′-CACGUAUUCCGACACGACUGGUAGCUG-5′(SEQ ID NO: 327)  AAT-1356 Target: 5′-GTGCATAAGGCTGTGCTGACCATCGAC-3′(SEQ ID NO: 525)  5′-CAUAAGGCUGUGCUGACCAUCGAcg-3′(SEQ ID NO: 130)  3′-ACGUAUUCCGACACGACUGGUAGCUGC-5′(SEQ ID NO: 328)  AAT-1357 Target: 5′-TGCATAAGGCTGTGCTGACCATCGACG-3′(SEQ ID NO: 526)  5′-AUAAGGCUGUGCUGACCAUCGACga-3′(SEQ ID NO: 131)  3′-CGUAUUCCGACACGACUGGUAGCUGCU-5′(SEQ ID NO: 329)  AAT-1358 Target: 5′-GCATAAGGCTGTGCTGACCATCGACGA-3′(SEQ ID NO: 527)  5′-UAAGGCUGUGCUGACCAUCGACGag-3′(SEQ ID NO: 132)  3′-GUAUUCCGACACGACUGGUAGCUGCUC-5′(SEQ ID NO: 330)  AAT-1359 Target: 5′-CATAAGGCTGTGCTGACCATCGACGAG-3′(SEQ ID NO: 528)  5′-AAGGCUGUGCUGACCAUCGACGAga-3′(SEQ ID NO: 133)  3′-UAUUCCGACACGACUGGUAGCUGCUCU-5′(SEQ ID NO: 331)  AAT-1360 Target: 5′-ATAAGGCTGTGCTGACCATCGACGAGA-3′(SEQ ID NO: 529)  5′-AGGCUGUGCUGACCAUCGACGAGaa-3′(SEQ ID NO: 134)  3′-AUUCCGACACGACUGGUAGCUGCUCUU-5′(SEQ ID NO: 332)  AAT-1361 Target: 5′-TAAGGCTGTGCTGACCATCGACGAGAA-3′(SEQ ID NO: 530)  5′-ACUGAAGCUGCUGGGGCCAUGUUtt-3′(SEQ ID NO: 135)  3′-CCUGACUUCGACGACCCCGGUACAAAA-5′(SEQ ID NO: 333)  AAT-1390 Target: 5′-GGACTGAAGCTGCTGGGGCCATGTTTT-3′(SEQ ID NO: 531)  5′-CUGAAGCUGCUGGGGCCAUGUUUtt-3′(SEQ ID NO: 136)  3′-CUGACUUCGACGACCCCGGUACAAAAA-5′(SEQ ID NO: 334)  AAT-1391 Target: 5′-GACTGAAGCTGCTGGGGCCATGTTTTT-3′(SEQ ID NO: 532)  5′-UGAAGCUGCUGGGGCCAUGUUUUta-3′(SEQ ID NO: 137)  3′-UGACUUCGACGACCCCGGUACAAAAAU-5′(SEQ ID NO: 335)  AAT-1392 Target: 5′-ACTGAAGCTGCTGGGGCCATGTTTTTA-3′(SEQ ID NO: 533)  5′-GAAGCUGCUGGGGCCAUGUUUUUag-3′(SEQ ID NO: 138)  3′-GACUUCGACGACCCCGGUACAAAAAUC-5′(SEQ ID NO: 336)  AAT-1393 Target: 5′-CTGAAGCTGCTGGGGCCATGTTTTTAG-3′(SEQ ID NO: 534)  5′-AAGCUGCUGGGGCCAUGUUUUUAga-3′(SEQ ID NO: 139)  3′-ACUUCGACGACCCCGGUACAAAAAUCU-5′(SEQ ID NO: 337)  AAT-1394 Target: 5′-TGAAGCTGCTGGGGCCATGTTTTTAGA-3′(SEQ ID NO: 535)  5′-AGCUGCUGGGGCCAUGUUUUUAGag-3′(SEQ ID NO: 140)  3′-CUUCGACGACCCCGGUACAAAAAUCUC-5′(SEQ ID NO: 338)  AAT-1395 Target: 5′-GAAGCTGCTGGGGCCATGTTTTTAGAG-3′(SEQ ID NO: 536)  5′-GCCAUGUUUUUAGAGGCCAUACCca-3′(SEQ ID NO: 141)  3′-CCCGGUACAAAAAUCUCCGGUAUGGGU-5′(SEQ ID NO: 339)  AAT-1405 Target: 5′-GGGCCATGTTTTTAGAGGCCATACCCA-3′(SEQ ID NO: 537)  5′-CCAUGUUUUUAGAGGCCAUACCCat-3′(SEQ ID NO: 142)  3′-CCGGUACAAAAAUCUCCGGUAUGGGUA-5′(SEQ ID NO: 340)  AAT-1406 Target: 5′-GGCCATGTTTTTAGAGGCCATACCCAT-3′(SEQ ID NO: 538)  5′-CAUGUUUUUAGAGGCCAUACCCAtg-3′(SEQ ID NO: 143)  3′-CGGUACAAAAAUCUCCGGUAUGGGUAC-5′(SEQ ID NO: 341)  AAT-1407 Target: 5′-GCCATGTTTTTAGAGGCCATACCCATG-3′(SEQ ID NO: 539)  5′-AUGUUUUUAGAGGCCAUACCCAUgt-3′(SEQ ID NO: 144)  3′-GGUACAAAAAUCUCCGGUAUGGGUACA-5′(SEQ ID NO: 342)  AAT-1408 Target: 5′-CCATGTTTTTAGAGGCCATACCCATGT-3′(SEQ ID NO: 540)  5′-UGUUUUUAGAGGCCAUACCCAUGtc-3′(SEQ ID NO: 145)  3′-GUACAAAAAUCUCCGGUAUGGGUACAG-5′(SEQ ID NO: 343)  AAT-1409 Target: 5′-CATGTTTTTAGAGGCCATACCCATGTC-3′(SEQ ID NO: 541)  5′-GUUUUUAGAGGCCAUACCCAUGUct-3′(SEQ ID NO: 146)  3′-UACAAAAAUCUCCGGUAUGGGUACAGA-5′(SEQ ID NO: 344)  AAT-1410 Target: 5′-ATGTTTTTAGAGGCCATACCCATGTCT-3′(SEQ ID NO: 542)  5′-UUUUUAGAGGCCAUACCCAUGUCta-3′(SEQ ID NO: 147)  3′-ACAAAAAUCUCCGGUAUGGGUACAGAU-5′(SEQ ID NO: 345)  AAT-1411 Target: 5′-TGTTTTTAGAGGCCATACCCATGTCTA-3′(SEQ ID NO: 543)  5′-UUUUAGAGGCCAUACCCAUGUCUat-3′(SEQ ID NO: 148)  3′-CAAAAAUCUCCGGUAUGGGUACAGAUA-5′(SEQ ID NO: 346)  AAT-1412 Target: 5′-GTTTTTAGAGGCCATACCCATGTCTAT-3′(SEQ ID NO: 544)  5′-UUUAGAGGCCAUACCCAUGUCUAtc-3′(SEQ ID NO: 149)  3′-AAAAAUCUCCGGUAUGGGUACAGAUAG-5′(SEQ ID NO: 347)  AAT-1413 Target: 5′-TTTTTAGAGGCCATACCCATGTCTATC-3′(SEQ ID NO: 545)  5′-UUAGAGGCCAUACCCAUGUCUAUcc-3′(SEQ ID NO: 150)  3′-AAAAUCUCCGGUAUGGGUACAGAUAGG-5′(SEQ ID NO: 348)  AAT-1414 Target: 5′-TTTTAGAGGCCATACCCATGTCTATCC-3′(SEQ ID NO: 546)  5′-UAGAGGCCAUACCCAUGUCUAUCcc-3′(SEQ ID NO: 151)  3′-AAAUCUCCGGUAUGGGUACAGAUAGGG-5′(SEQ ID NO: 349)  AAT-1415 Target: 5′-TTTAGAGGCCATACCCATGTCTATCCC-3′(SEQ ID NO: 547)  5′-AGAGGCCAUACCCAUGUCUAUCCcc-3′(SEQ ID NO: 152)  3′-AAUCUCCGGUAUGGGUACAGAUAGGGG-5′(SEQ ID NO: 350)  AAT-1416 Target: 5′-TTAGAGGCCATACCCATGTCTATCCCC-3′(SEQ ID NO: 548)  5′-GUUCAACAAACCCUUUGUCUUCUta-3′(SEQ ID NO: 153)  3′-UUCAAGUUGUUUGGGAAACAGAAGAAU-5′(SEQ ID NO: 351)  AAT-1452 Target: 5′-AAGTTCAACAAACCCTTTGTCTTCTTA-3′(SEQ ID NO: 549)  5′-UUCAACAAACCCUUUGUCUUCUUaa-3′(SEQ ID NO: 154)  3′-UCAAGUUGUUUGGGAAACAGAAGAAUU-5′(SEQ ID NO: 352)  AAT-1453 Target: 5′-AGTTCAACAAACCCTTTGTCTTCTTAA-3′(SEQ ID NO: 550)  5′-UCAACAAACCCUUUGUCUUCUUAat-3′(SEQ ID NO: 155)  3′-CAAGUUGUUUGGGAAACAGAAGAAUUA-5′(SEQ ID NO: 353)  AAT-1454 Target: 5′-GTTCAACAAACCCTTTGTCTTCTTAAT-3′(SEQ ID NO: 551)  5′-CAACAAACCCUUUGUCUUCUUAAtg-3′(SEQ ID NO: 156)  3′-AAGUUGUUUGGGAAACAGAAGAAUUAC-5′(SEQ ID NO: 354)  AAT-1455 Target: 5′-TTCAACAAACCCTTTGTCTTCTTAATG-3′(SEQ ID NO: 552)  5′-AACAAACCCUUUGUCUUCUUAAUga-3′(SEQ ID NO: 157)  3′-AGUUGUUUGGGAAACAGAAGAAUUACU-5′(SEQ ID NO: 355)  AAT-1456 Target: 5′-TCAACAAACCCTTTGTCTTCTTAATGA-3′(SEQ ID NO: 553)  5′-ACAAACCCUUUGUCUUCUUAAUGat-3′(SEQ ID NO: 158)  3′-GUUGUUUGGGAAACAGAAGAAUUACUA-5′(SEQ ID NO: 356)  AAT-1457 Target: 5′-CAACAAACCCTTTGTCTTCTTAATGAT-3′(SEQ ID NO: 554)  5′-CAAACCCUUUGUCUUCUUAAUGAtt-3′(SEQ ID NO: 159)  3′-UUGUUUGGGAAACAGAAGAAUUACUAA-5′(SEQ ID NO: 357)  AAT-1458 Target: 5′-AACAAACCCTTTGTCTTCTTAATGATT-3′(SEQ ID NO: 555)  5′-AAACCCUUUGUCUUCUUAAUGAUtg-3′(SEQ ID NO: 160)  3′-UGUUUGGGAAACAGAAGAAUUACUAAC-5′(SEQ ID NO: 358)  AAT-1459 Target: 5′-ACAAACCCTTTGTCTTCTTAATGATTG-3′(SEQ ID NO: 556)  5′-AACCCUUUGUCUUCUUAAUGAUUga-3′(SEQ ID NO: 161)  3′-GUUUGGGAAACAGAAGAAUUACUAACU-5′(SEQ ID NO: 359)  AAT-1460 Target: 5′-CAAACCCTTTGTCTTCTTAATGATTGA-3′(SEQ ID NO: 557)  5′-AAUACCAAGUCUCCCCUCUUCAUgg-3′(SEQ ID NO: 162)  3′-UUUUAUGGUUCAGAGGGGAGAAGUACC-5′(SEQ ID NO: 360)  AAT-1489 Target: 5′-AAAATACCAAGTCTCCCCTCTTCATGG-3′(SEQ ID NO: 558)  5′-AUACCAAGUCUCCCCUCUUCAUGgg-3′(SEQ ID NO: 163)  3′-UUUAUGGUUCAGAGGGGAGAAGUACCC-5′(SEQ ID NO: 361)  AAT-1490 Target: 5′-AAATACCAAGTCTCCCCTCTTCATGGG-3′(SEQ ID NO: 559)  5′-UACCAAGUCUCCCCUCUUCAUGGga-3′(SEQ ID NO: 164)  3′-UUAUGGUUCAGAGGGGAGAAGUACCCU-5′(SEQ ID NO: 362)  AAT-1491 Target: 5′-AATACCAAGTCTCCCCTCTTCATGGGA-3′(SEQ ID NO: 560)  5′-ACCAAGUCUCCCCUCUUCAUGGGaa-3′(SEQ ID NO: 165)  3′-UAUGGUUCAGAGGGGAGAAGUACCCUU-5′(SEQ ID NO: 363)  AAT-1492 Target: 5′-ATACCAAGTCTCCCCTCTTCATGGGAA-3′(SEQ ID NO: 561)  5′-CCAAGUCUCCCCUCUUCAUGGGAaa-3′(SEQ ID NO: 166)  3′-AUGGUUCAGAGGGGAGAAGUACCCUUU-5′(SEQ ID NO: 364)  AAT-1493 Target: 5′-TACCAAGTCTCCCCTCTTCATGGGAAA-3′(SEQ ID NO: 562)  5′-CAAGUCUCCCCUCUUCAUGGGAAaa-3′(SEQ ID NO: 167)  3′-UGGUUCAGAGGGGAGAAGUACCCUUUU-5′(SEQ ID NO: 365)  AAT-1494 Target: 5′-ACCAAGTCTCCCCTCTTCATGGGAAAA-3′(SEQ ID NO: 563)  5′-AAGUCUCCCCUCUUCAUGGGAAAag-3′(SEQ ID NO: 168)  3′-GGUUCAGAGGGGAGAAGUACCCUUUUC-5′(SEQ ID NO: 366)  AAT-1495 Target: 5′-CCAAGTCTCCCCTCTTCATGGGAAAAG-3′(SEQ ID NO: 564)  5′-AGUCUCCCCUCUUCAUGGGAAAAgt-3′(SEQ ID NO: 169)  3′-GUUCAGAGGGGAGAAGUACCCUUUUCA-5′(SEQ ID NO: 367)  AAT-1496 Target: 5′-CAAGTCTCCCCTCTTCATGGGAAAAGT-3′(SEQ ID NO: 565)  5′-GUCUCCCCUCUUCAUGGGAAAAGtg-3′(SEQ ID NO: 170)  3′-UUCAGAGGGGAGAAGUACCCUUUUCAC-5′(SEQ ID NO: 368)  AAT-1497 Target: 5′-AAGTCTCCCCTCTTCATGGGAAAAGTG-3′(SEQ ID NO: 566)  5′-CUCCCCUCUUCAUGGGAAAAGUGgt-3′(SEQ ID NO: 171)  3′-CAGAGGGGAGAAGUACCCUUUUCACCA-5′(SEQ ID NO: 369)  AAT-1499 Target: 5′-GTCTCCCCTCTTCATGGGAAAAGTGGT-3′(SEQ ID NO: 567)  5′-CCCCUCUUCAUGGGAAAAGUGGUga-3′(SEQ ID NO: 172)  3′-GAGGGGAGAAGUACCCUUUUCACCACU-5′(SEQ ID NO: 370)  AAT-1501 Target: 5′-CTCCCCTCTTCATGGGAAAAGTGGTGA-3′(SEQ ID NO: 568)  5′-CCCUCUUCAUGGGAAAAGUGGUGaa-3′(SEQ ID NO: 173)  3′-AGGGGAGAAGUACCCUUUUCACCACUU-5′(SEQ ID NO: 371)  AAT-1502 Target: 5′-TCCCCTCTTCATGGGAAAAGTGGTGAA-3′(SEQ ID NO: 569)  5′-CCUCUUCAUGGGAAAAGUGGUGAat-3′(SEQ ID NO: 174)  3′-GGGGAGAAGUACCCUUUUCACCACUUA-5′(SEQ ID NO: 372)  AAT-1503 Target: 5′-CCCCTCTTCATGGGAAAAGTGGTGAAT-3′(SEQ ID NO: 570)  5′-CUCUUCAUGGGAAAAGUGGUGAAtc-3′(SEQ ID NO: 175)  3′-GGGAGAAGUACCCUUUUCACCACUUAG-5′(SEQ ID NO: 373)  AAT-1504 Target: 5′-CCCTCTTCATGGGAAAAGTGGTGAATC-3′(SEQ ID NO: 571)  5′-UCUUCAUGGGAAAAGUGGUGAAUcc-3′(SEQ ID NO: 176)  3′-GGAGAAGUACCCUUUUCACCACUUAGG-5′(SEQ ID NO: 374)  AAT-1505 Target: 5′-CCTCTTCATGGGAAAAGTGGTGAATCC-3′(SEQ ID NO: 572)  5′-CUUCAUGGGAAAAGUGGUGAAUCcc-3′(SEQ ID NO: 177)  3′-GAGAAGUACCCUUUUCACCACUUAGGG-5′(SEQ ID NO: 375)  AAT-1506 Target: 5′-CTCTTCATGGGAAAAGTGGTGAATCCC-3′(SEQ ID NO: 573)  5′-UUCAUGGGAAAAGUGGUGAAUCCca-3′(SEQ ID NO: 178)  3′-AGAAGUACCCUUUUCACCACUUAGGGU-5′(SEQ ID NO: 376)  AAT-1507 Target: 5′-TCTTCATGGGAAAAGTGGTGAATCCCA-3′(SEQ ID NO: 574)  5′-UCAUGGGAAAAGUGGUGAAUCCCac-3′(SEQ ID NO: 179)  3′-GAAGUACCCUUUUCACCACUUAGGGUG-5′(SEQ ID NO: 377)  AAT-1508 Target: 5′-CTTCATGGGAAAAGTGGTGAATCCCAC-3′(SEQ ID NO: 575)  5′-CAUGGGAAAAGUGGUGAAUCCCAcc-3′(SEQ ID NO: 180)  3′-AAGUACCCUUUUCACCACUUAGGGUGG-5′(SEQ ID NO: 378)  AAT-1509 Target: 5′-TTCATGGGAAAAGTGGTGAATCCCACC-3′(SEQ ID NO: 576)  5′-AUGGGAAAAGUGGUGAAUCCCACcc-3′(SEQ ID NO: 181)  3′-AGUACCCUUUUCACCACUUAGGGUGGG-5′(SEQ ID NO: 379)  AAT-1510 Target: 5′-TCATGGGAAAAGTGGTGAATCCCACCC-3′(SEQ ID NO: 577)  5′-UGGGAAAAGUGGUGAAUCCCACCca-3′(SEQ ID NO: 182)  3′-GUACCCUUUUCACCACUUAGGGUGGGU-5′(SEQ ID NO: 380)  AAT-1511 Target: 5′-CATGGGAAAAGTGGTGAATCCCACCCA-3′(SEQ ID NO: 578)  5′-GGGAAAAGUGGUGAAUCCCACCCaa-3′(SEQ ID NO: 183)  3′-UACCCUUUUCACCACUUAGGGUGGGUU-5′(SEQ ID NO: 381)  AAT-1512 Target: 5′-ATGGGAAAAGTGGTGAATCCCACCCAA-3′(SEQ ID NO: 579)  5′-GGAAAAGUGGUGAAUCCCACCCAaa-3′(SEQ ID NO: 184)  3′-ACCCUUUUCACCACUUAGGGUGGGUUU-5′(SEQ ID NO: 382)  AAT-1513 Target: 5′-TGGGAAAAGTGGTGAATCCCACCCAAA-3′(SEQ ID NO: 580)  5′-GAAAAGUGGUGAAUCCCACCCAAaa-3′(SEQ ID NO: 185)  3′-CCCUUUUCACCACUUAGGGUGGGUUUU-5′(SEQ ID NO: 383)  AAT-1514 Target: 5′-GGGAAAAGTGGTGAATCCCACCCAAAA-3′(SEQ ID NO: 581)  5′-AAAAGUGGUGAAUCCCACCCAAAaa-3′(SEQ ID NO: 186)  3′-CCUUUUCACCACUUAGGGUGGGUUUUU-5′(SEQ ID NO: 384)  AAT-1515 Target: 5′-GGAAAAGTGGTGAATCCCACCCAAAAA-3′(SEQ ID NO: 582)  5′-AAAGUGGUGAAUCCCACCCAAAAat-3′(SEQ ID NO: 187)  3′-CUUUUCACCACUUAGGGUGGGUUUUUA-5′(SEQ ID NO: 385)  AAT-1516 Target: 5′-GAAAAGTGGTGAATCCCACCCAAAAAT-3′(SEQ ID NO: 583)  5′-AAGUGGUGAAUCCCACCCAAAAAta-3′(SEQ ID NO: 188)  3′-UUUUCACCACUUAGGGUGGGUUUUUAU-5′(SEQ ID NO: 386)  AAT-1517 Target: 5′-AAAAGTGGTGAATCCCACCCAAAAATA-3′(SEQ ID NO: 584)  5′-CGAUAGUUCAAAAUGGUGAAAUUag-3′(SEQ ID NO: 189)  3′-AAGCUAUCAAGUUUUACCACUUUAAUC-5′(SEQ ID NO: 387)  AAT-2872 Target: 5′-TTCGATAGTTCAAAATGGTGAAATTAG-3′(SEQ ID NO: 585)  5′-CAAAAUGGUGAAAUUAGCAAUUCta-3′(SEQ ID NO: 190)  3′-AAGUUUUACCACUUUAAUCGUUAAGAU-5′(SEQ ID NO: 388)  AAT-2880 Target: 5′-TTCAAAATGGTGAAATTAGCAATTCTA-3′(SEQ ID NO: 586)  5′-UUGGUAUGAUGUUCAAGUUAGAUaa-3′(SEQ ID NO: 191)  3′-UCAACCAUACUACAAGUUCAAUCUAUU-5′(SEQ ID NO: 389)  AAT-3167 Target: 5′-AGTTGGTATGATGTTCAAGTTAGATAA-3′(SEQ ID NO: 587)  5′-GGUAUGAUGUUCAAGUUAGAUAAca-3′(SEQ ID NO: 192)  3′-AACCAUACUACAAGUUCAAUCUAUUGU-5′(SEQ ID NO: 390)  AAT-3169 Target: 5′-TTGGTATGATGTTCAAGTTAGATAACA-3′(SEQ ID NO: 588)  5′-GUAUGAUGUUCAAGUUAGAUAACaa-3′(SEQ ID NO: 193)  3′-ACCAUACUACAAGUUCAAUCUAUUGUU-5′(SEQ ID NO: 391)  AAT-3170 Target: 5′-TGGTATGATGTTCAAGTTAGATAACAA-3′(SEQ ID NO: 589)  5′-AUGAUGUUCAAGUUAGAUAACAAaa-3′(SEQ ID NO: 194)  3′-CAUACUACAAGUUCAAUCUAUUGUUUU-5′(SEQ ID NO: 392)  AAT-3172 Target: 5′-GTATGATGTTCAAGTTAGATAACAAAA-3′(SEQ ID NO: 590)  5′-AUGUUCAAGUUAGAUAACAAAAUgt-3′(SEQ ID NO: 195)  3′-ACUACAAGUUCAAUCUAUUGUUUUACA-5′(SEQ ID NO: 393)  AAT-3175 Target: 5′-TGATGTTCAAGTTAGATAACAAAATGT-3′(SEQ ID NO: 591)  5′-CAAGUUAGAUAACAAAAUGUUUAta-3′(SEQ ID NO: 196)  3′-AAGUUCAAUCUAUUGUUUUACAAAUAU-5′(SEQ ID NO: 394)  AAT-3180 Target: 5′-TTCAAGTTAGATAACAAAATGTTTATA-3′(SEQ ID NO: 592)  5′-AAGUUAGAUAACAAAAUGUUUAUac-3′(SEQ ID NO: 197)  3′-AGUUCAAUCUAUUGUUUUACAAAUAUG-5′(SEQ ID NO: 395)  AAT-3181 Target: 5′-TCAAGTTAGATAACAAAATGTTTATAC-3′(SEQ ID NO: 593)  5′-AGUUAGAUAACAAAAUGUUUAUAcc-3′(SEQ ID NO: 198)  3′-GUUCAAUCUAUUGUUUUACAAAUAUGG-5′(SEQ ID NO: 396)  AAT-3182 Target: 5′-CAAGTTAGATAACAAAATGTTTATACC-3′(SEQ ID NO: 594)

TABLE 3 Selected Human Anti-α-1 antitrypsin DsiRNAs, Unmodified Duplexes  (Asymmetrics) 5′-CAUCCCACCAUGAUCAGGAUCACCC-3′(SEQ ID NO: 595)  3′-AUGUAGGGUGGUACUAGUCCUAGUGGG-5′(SEQ ID NO: 199)  AAT-395 Target: 5′-TACATCCCACCATGATCAGGATCACCC-3′(SEQ ID NO: 397)  5′-CAGCUGGCACACCAGUCCAACAGCA-3′(SEQ ID NO: 596)  3′-CGGUCGACCGUGUGGUCAGGUUGUCGU-5′(SEQ ID NO: 200)  AAT-475 Target: 5′-GCCAGCTGGCACACCAGTCCAACAGCA-3′(SEQ ID NO: 398)  5′-GCUGGCACACCAGUCCAACAGCACC-3′(SEQ ID NO: 597)  3′-GUCGACCGUGUGGUCAGGUUGUCGUGG-5′(SEQ ID NO: 201)  AAT-477 Target: 5′-CAGCTGGCACACCAGTCCAACAGCACC-3′(SEQ ID NO: 399)  5′-GGCACACCAGUCCAACAGCACCAAU-3′(SEQ ID NO: 598)  3′-GACCGUGUGGUCAGGUUGUCGUGGUUA-5′(SEQ ID NO: 202)  AAT-480 Target: 5′-CTGGCACACCAGTCCAACAGCACCAAT-3′(SEQ ID NO: 400)  5′-GCACACCAGUCCAACAGCACCAAUA-3′(SEQ ID NO: 599)  3′-ACCGUGUGGUCAGGUUGUCGUGGUUAU-5′(SEQ ID NO: 203)  AAT-481 Target: 5′-TGGCACACCAGTCCAACAGCACCAATA-3′(SEQ ID NO: 401)  5′-CACACCAGUCCAACAGCACCAAUAU-3′(SEQ ID NO: 600)  3′-CCGUGUGGUCAGGUUGUCGUGGUUAUA-5′(SEQ ID NO: 204)  AAT-482 Target: 5′-GGCACACCAGTCCAACAGCACCAATAT-3′(SEQ ID NO: 402)  5′-ACACCAGUCCAACAGCACCAAUAUC-3′(SEQ ID NO: 601)  3′-CGUGUGGUCAGGUUGUCGUGGUUAUAG-5′(SEQ ID NO: 205)  AAT-483 Target: 5′-GCACACCAGTCCAACAGCACCAATATC-3′(SEQ ID NO: 403)  5′-CACCAGUCCAACAGCACCAAUAUCU-3′(SEQ ID NO: 602)  3′-GUGUGGUCAGGUUGUCGUGGUUAUAGA-5′(SEQ ID NO: 206)  AAT-484 Target: 5′-CACACCAGTCCAACAGCACCAATATCT-3′(SEQ ID NO: 404)  5′-CCAAUAUCUUCUUCUCCCCAGUGAG-3′(SEQ ID NO: 603)  3′-GUGGUUAUAGAAGAAGAGGGGUCACUC-5′(SEQ ID NO: 207)  AAT-500 Target: 5′-CACCAATATCTTCTTCTCCCCAGTGAG-3′(SEQ ID NO: 405)  5′-CAAUAUCUUCUUCUCCCCAGUGAGC-3′(SEQ ID NO: 604)  3′-UGGUUAUAGAAGAAGAGGGGUCACUCG-5′(SEQ ID NO: 208)  AAT-501 Target: 5′-ACCAATATCTTCTTCTCCCCAGTGAGC-3′(SEQ ID NO: 406)  5′-AAUAUCUUCUUCUCCCCAGUGAGCA-3′(SEQ ID NO: 605)  3′-GGUUAUAGAAGAAGAGGGGUCACUCGU-5′(SEQ ID NO: 209)  AAT-502 Target: 5′-CCAATATCTTCTTCTCCCCAGTGAGCA-3′(SEQ ID NO: 407)  5′-AUAUCUUCUUCUCCCCAGUGAGCAU-3′(SEQ ID NO: 606)  3′-GUUAUAGAAGAAGAGGGGUCACUCGUA-5′(SEQ ID NO: 210)  AAT-503 Target: 5′-CAATATCTTCTTCTCCCCAGTGAGCAT-3′(SEQ ID NO: 408)  5′-UAUCUUCUUCUCCCCAGUGAGCAUC-3′(SEQ ID NO: 607)  3′-UUAUAGAAGAAGAGGGGUCACUCGUAG-5′(SEQ ID NO: 211)  AAT-504 Target: 5′-AATATCTTCTTCTCCCCAGTGAGCATC-3′(SEQ ID NO: 409)  5′-AUCUUCUUCUCCCCAGUGAGCAUCG-3′(SEQ ID NO: 608)  3′-UAUAGAAGAAGAGGGGUCACUCGUAGC-5′(SEQ ID NO: 212)  AAT-505 Target: 5′-ATATCTTCTTCTCCCCAGTGAGCATCG-3′(SEQ ID NO: 410)  5′-UCUUCUUCUCCCCAGUGAGCAUCGC-3′(SEQ ID NO: 609)  3′-AUAGAAGAAGAGGGGUCACUCGUAGCG-5′(SEQ ID NO: 213)  AAT-506 Target: 5′-TATCTTCTTCTCCCCAGTGAGCATCGC-3′(SEQ ID NO: 411)  5′-CUUCUUCUCCCCAGUGAGCAUCGCU-3′(SEQ ID NO: 610)  3′-UAGAAGAAGAGGGGUCACUCGUAGCGA-5′(SEQ ID NO: 214)  AAT-507 Target: 5′-ATCTTCTTCTCCCCAGTGAGCATCGCT-3′(SEQ ID NO: 412)  5′-UUCUUCUCCCCAGUGAGCAUCGCUA-3′(SEQ ID NO: 611)  3′-AGAAGAAGAGGGGUCACUCGUAGCGAU-5′(SEQ ID NO: 215)  AAT-508 Target: 5′-TCTTCTTCTCCCCAGTGAGCATCGCTA-3′(SEQ ID NO: 413)  5′-UCUUCUCCCCAGUGAGCAUCGCUAC-3′(SEQ ID NO: 612)  3′-GAAGAAGAGGGGUCACUCGUAGCGAUG-5′(SEQ ID NO: 216)  AAT-509 Target: 5′-CTTCTTCTCCCCAGTGAGCATCGCTAC-3′(SEQ ID NO: 414)  5′-CUUCUCCCCAGUGAGCAUCGCUACA-3′(SEQ ID NO: 613)  3′-AAGAAGAGGGGUCACUCGUAGCGAUGU-5′(SEQ ID NO: 217)  AAT-510 Target: 5′-TTCTTCTCCCCAGTGAGCATCGCTACA-3′(SEQ ID NO: 415)  5′-UCUCCCCAGUGAGCAUCGCUACAGC-3′(SEQ ID NO: 614)  3′-GAAGAGGGGUCACUCGUAGCGAUGUCG-5′(SEQ ID NO: 218)  AAT-512 Target: 5′-CTTCTCCCCAGTGAGCATCGCTACAGC-3′(SEQ ID NO: 416)  5′-CUCCCCAGUGAGCAUCGCUACAGCC-3′(SEQ ID NO: 615)  3′-AAGAGGGGUCACUCGUAGCGAUGUCGG-5′(SEQ ID NO: 219)  AAT-513 Target: 5′-TTCTCCCCAGTGAGCATCGCTACAGCC-3′(SEQ ID NO: 417)  5′-CCCCAGUGAGCAUCGCUACAGCCUU-3′(SEQ ID NO: 616)  3′-GAGGGGUCACUCGUAGCGAUGUCGGAA-5′(SEQ ID NO: 220)  AAT-515 Target: 5′-CTCCCCAGTGAGCATCGCTACAGCCTT-3′(SEQ ID NO: 418)  5′-ACAGCCUUUGCAAUGCUCUCCCUGG-3′(SEQ ID NO: 617)  3′-GAUGUCGGAAACGUUACGAGAGGGACC-5′(SEQ ID NO: 221)  AAT-532 Target: 5′-CTACAGCCTTTGCAATGCTCTCCCTGG-3′(SEQ ID NO: 419)  5′-UGCAAUGCUCUCCCUGGGGACCAAG-3′(SEQ ID NO: 618)  3′-AAACGUUACGAGAGGGACCCCUGGUUC-5′(SEQ ID NO: 222)  AAT-540 Target: 5′-TTTGCAATGCTCTCCCTGGGGACCAAG-3′(SEQ ID NO: 420)  5′-AAAUCCUGGAGGGCCUGAAUUUCAA-3′(SEQ ID NO: 619)  3′-ACUUUAGGACCUCCCGGACUUAAAGUU-5′(SEQ ID NO: 223)  AAT-581 Target: 5′-TGAAATCCTGGAGGGCCTGAATTTCAA-3′(SEQ ID NO: 421)  5′-AAUCCUGGAGGGCCUGAAUUUCAAC-3′(SEQ ID NO: 620)  3′-CUUUAGGACCUCCCGGACUUAAAGUUG-5′(SEQ ID NO: 224)  AAT-582 Target: 5′-GAAATCCTGGAGGGCCTGAATTTCAAC-3′(SEQ ID NO: 422)  5′-AUCCUGGAGGGCCUGAAUUUCAACC-3′(SEQ ID NO: 621)  3′-UUUAGGACCUCCCGGACUUAAAGUUGG-5′(SEQ ID NO: 225)  AAT-583 Target: 5′-AAATCCTGGAGGGCCTGAATTTCAACC-3′(SEQ ID NO: 423)  5′-CCUGGAGGGCCUGAAUUUCAACCUC-3′(SEQ ID NO: 622)  3′-UAGGACCUCCCGGACUUAAAGUUGGAG-5′(SEQ ID NO: 226)  AAT-585 Target: 5′-ATCCTGGAGGGCCTGAATTTCAACCTC-3′(SEQ ID NO: 424)  5′-CUGGAGGGCCUGAAUUUCAACCUCA-3′(SEQ ID NO: 623)  3′-AGGACCUCCCGGACUUAAAGUUGGAGU-5′(SEQ ID NO: 227)  AAT-586 Target: 5′-TCCTGGAGGGCCTGAATTTCAACCTCA-3′(SEQ ID NO: 425)  5′-UGGAGGGCCUGAAUUUCAACCUCAC-3′(SEQ ID NO: 624)  3′-GGACCUCCCGGACUUAAAGUUGGAGUG-5′(SEQ ID NO: 228)  AAT-587 Target: 5′-CCTGGAGGGCCTGAATTTCAACCTCAC-3′(SEQ ID NO: 426)  5′-CAUGAAGGCUUCCAGGAACUCCUCC-3′(SEQ ID NO: 625)  3′-AGGUACUUCCGAAGGUCCUUGAGGAGG-5′(SEQ ID NO: 229)  AAT-634 Target: 5′-TCCATGAAGGCTTCCAGGAACTCCTCC-3′(SEQ ID NO: 427)  5′-GAAGGCUUCCAGGAACUCCUCCGUA-3′(SEQ ID NO: 626)  3′-UACUUCCGAAGGUCCUUGAGGAGGCAU-5′(SEQ ID NO: 230)  AAT-637 Target: 5′-ATGAAGGCTTCCAGGAACTCCTCCGTA-3′(SEQ ID NO: 428)  5′-AAGGCUUCCAGGAACUCCUCCGUAC-3′(SEQ ID NO: 627)  3′-ACUUCCGAAGGUCCUUGAGGAGGCAUG-5′(SEQ ID NO: 231)  AAT-638 Target: 5′-TGAAGGCTTCCAGGAACTCCTCCGTAC-3′(SEQ ID NO: 429)  5′-AGCCAGACAGCCAGCUCCAGCUGAC-3′(SEQ ID NO: 628)  3′-GGUCGGUCUGUCGGUCGAGGUCGACUG-5′(SEQ ID NO: 232)  AAT-671 Target: 5′-CCAGCCAGACAGCCAGCTCCAGCTGAC-3′(SEQ ID NO: 430)  5′-CCAGACAGCCAGCUCCAGCUGACCA-3′(SEQ ID NO: 629)  3′-UCGGUCUGUCGGUCGAGGUCGACUGGU-5′(SEQ ID NO: 233)  AAT-673 Target: 5′-AGCCAGACAGCCAGCTCCAGCTGACCA-3′(SEQ ID NO: 431)  5′-AGACAGCCAGCUCCAGCUGACCACC-3′(SEQ ID NO: 630)  3′-GGUCUGUCGGUCGAGGUCGACUGGUGG-5′(SEQ ID NO: 234)  AAT-675 Target: 5′-CCAGACAGCCAGCTCCAGCTGACCACC-3′(SEQ ID NO: 432)  5′-GACAGCCAGCUCCAGCUGACCACCG-3′(SEQ ID NO: 631)  3′-GUCUGUCGGUCGAGGUCGACUGGUGGC-5′(SEQ ID NO: 235)  AAT-676 Target: 5′-CAGACAGCCAGCTCCAGCTGACCACCG-3′(SEQ ID NO: 433)  5′-UAGUGGAUAAGUUUUUGGAGGAUGU-3′(SEQ ID NO: 632)  3′-CGAUCACCUAUUCAAAAACCUCCUACA-5′(SEQ ID NO: 236)  AAT-734 Target: 5′-GCTAGTGGATAAGTTTTTGGAGGATGT-3′(SEQ ID NO: 434)  5′-AGUGGAUAAGUUUUUGGAGGAUGUU-3′(SEQ ID NO: 633)  3′-GAUCACCUAUUCAAAAACCUCCUACAA-5′(SEQ ID NO: 237)  AAT-735 Target: 5′-CTAGTGGATAAGTTTTTGGAGGATGTT-3′(SEQ ID NO: 435)  5′-GUGGAUAAGUUUUUGGAGGAUGUUA-3′(SEQ ID NO: 634)  3′-AUCACCUAUUCAAAAACCUCCUACAAU-5′(SEQ ID NO: 238)  AAT-736 Target: 5′-TAGTGGATAAGTTTTTGGAGGATGTTA-3′(SEQ ID NO: 436)  5′-UGGAUAAGUUUUUGGAGGAUGUUAA-3′(SEQ ID NO: 635)  3′-UCACCUAUUCAAAAACCUCCUACAAUU-5′(SEQ ID NO: 239)  AAT-737 Target: 5′-AGTGGATAAGTTTTTGGAGGATGTTAA-3′(SEQ ID NO: 437)  5′-GGAUAAGUUUUUGGAGGAUGUUAAA-3′(SEQ ID NO: 636)  3′-CACCUAUUCAAAAACCUCCUACAAUUU-5′(SEQ ID NO: 240)  AAT-738 Target: 5′-GTGGATAAGTTTTTGGAGGATGTTAAA-3′(SEQ ID NO: 438)  5′-GAUAAGUUUUUGGAGGAUGUUAAAA-3′(SEQ ID NO: 637)  3′-ACCUAUUCAAAAACCUCCUACAAUUUU-5′(SEQ ID NO: 241)  AAT-739 Target: 5′-TGGATAAGTTTTTGGAGGATGTTAAAA-3′(SEQ ID NO: 439)  5′-AUAAGUUUUUGGAGGAUGUUAAAAA-3′(SEQ ID NO: 638)  3′-CCUAUUCAAAAACCUCCUACAAUUUUU-5′(SEQ ID NO: 242)  AAT-740 Target: 5′-GGATAAGTTTTTGGAGGATGTTAAAAA-3′(SEQ ID NO: 440)  5′-UGUACCACUCAGAAGCCUUCACUGU-3′(SEQ ID NO: 639)  3′-CAACAUGGUGAGUCUUCGGAAGUGACA-5′(SEQ ID NO: 243)  AAT-767 Target: 5′-GTTGTACCACTCAGAAGCCTTCACTGT-3′(SEQ ID NO: 441)  5′-GUACCACUCAGAAGCCUUCACUGUC-3′(SEQ ID NO: 640)  3′-AACAUGGUGAGUCUUCGGAAGUGACAG-5′(SEQ ID NO: 244)  AAT-768 Target: 5′-TTGTACCACTCAGAAGCCTTCACTGTC-3′(SEQ ID NO: 442)  5′-ACUCAAGGGAAAAUUGUGGAUUUGG-3′(SEQ ID NO: 641)  3′-CAUGAGUUCCCUUUUAACACCUAAACC-5′(SEQ ID NO: 245)  AAT-850 Target: 5′-GTACTCAAGGGAAAATTGTGGATTTGG-3′(SEQ ID NO: 443)  5′-CUCAAGGGAAAAUUGUGGAUUUGGU-3′(SEQ ID NO: 642)  3′-AUGAGUUCCCUUUUAACACCUAAACCA-5′(SEQ ID NO: 246)  AAT-851 Target: 5′-TACTCAAGGGAAAATTGTGGATTTGGT-3′(SEQ ID NO: 444)  5′-UCAAGGGAAAAUUGUGGAUUUGGUC-3′(SEQ ID NO: 643)  3′-UGAGUUCCCUUUUAACACCUAAACCAG-5′(SEQ ID NO: 247)  AAT-852 Target: 5′-ACTCAAGGGAAAATTGTGGATTTGGTC-3′(SEQ ID NO: 445)  5′-CAAGGGAAAAUUGUGGAUUUGGUCA-3′(SEQ ID NO: 644)  3′-GAGUUCCCUUUUAACACCUAAACCAGU-5′(SEQ ID NO: 248)  AAT-853 Target: 5′-CTCAAGGGAAAATTGTGGATTTGGTCA-3′(SEQ ID NO: 446)  5′-AAGGGAAAAUUGUGGAUUUGGUCAA-3′(SEQ ID NO: 645)  3′-AGUUCCCUUUUAACACCUAAACCAGUU-5′(SEQ ID NO: 249)  AAT-854 Target: 5′-TCAAGGGAAAATTGTGGATTTGGTCAA-3′(SEQ ID NO: 447)  5′-AGGGAAAAUUGUGGAUUUGGUCAAG-3′(SEQ ID NO: 646)  3′-GUUCCCUUUUAACACCUAAACCAGUUC-5′(SEQ ID NO: 250)  AAT-855 Target: 5′-CAAGGGAAAATTGTGGATTTGGTCAAG-3′(SEQ ID NO: 448)  5′-GGGAAAAUUGUGGAUUUGGUCAAGG-3′(SEQ ID NO: 647)  3′-UUCCCUUUUAACACCUAAACCAGUUCC-5′(SEQ ID NO: 251)  AAT-856 Target: 5′-AAGGGAAAATTGTGGATTTGGTCAAGG-3′(SEQ ID NO: 449)  5′-GGAAAAUUGUGGAUUUGGUCAAGGA-3′(SEQ ID NO: 648)  3′-UCCCUUUUAACACCUAAACCAGUUCCU-5′(SEQ ID NO: 252)  AAT-857 Target: 5′-AGGGAAAATTGTGGATTTGGTCAAGGA-3′(SEQ ID NO: 450)  5′-GAAAAUUGUGGAUUUGGUCAAGGAG-3′(SEQ ID NO: 649)  3′-CCCUUUUAACACCUAAACCAGUUCCUC-5′(SEQ ID NO: 253)  AAT-858 Target: 5′-GGGAAAATTGTGGATTTGGTCAAGGAG-3′(SEQ ID NO: 451)  5′-AAAAUUGUGGAUUUGGUCAAGGAGC-3′(SEQ ID NO: 650)  3′-CCUUUUAACACCUAAACCAGUUCCUCG-5′(SEQ ID NO: 254)  AAT-859 Target: 5′-GGAAAATTGTGGATTTGGTCAAGGAGC-3′(SEQ ID NO: 452)  5′-AAAUUGUGGAUUUGGUCAAGGAGCU-3′(SEQ ID NO: 651)  3′-CUUUUAACACCUAAACCAGUUCCUCGA-5′(SEQ ID NO: 255)  AAT-860 Target: 5′-GAAAATTGTGGATTTGGTCAAGGAGCT-3′(SEQ ID NO: 453)  5′-AAUUGUGGAUUUGGUCAAGGAGCUU-3′(SEQ ID NO: 652)  3′-UUUUAACACCUAAACCAGUUCCUCGAA-5′(SEQ ID NO: 256)  AAT-861 Target: 5′-AAAATTGTGGATTTGGTCAAGGAGCTT-3′(SEQ ID NO: 454)  5′-AUUGUGGAUUUGGUCAAGGAGCUUG-3′(SEQ ID NO: 653)  3′-UUUAACACCUAAACCAGUUCCUCGAAC-5′(SEQ ID NO: 257)  AAT-862 Target: 5′-AAATTGTGGATTTGGTCAAGGAGCTTG-3′(SEQ ID NO: 455)  5′-UUGUGGAUUUGGUCAAGGAGCUUGA-3′(SEQ ID NO: 654)  3′-UUAACACCUAAACCAGUUCCUCGAACU-5′(SEQ ID NO: 258)  AAT-863 Target: 5′-AATTGTGGATTTGGTCAAGGAGCTTGA-3′(SEQ ID NO: 456)  5′-UGUGGAUUUGGUCAAGGAGCUUGAC-3′(SEQ ID NO: 655)  3′-UAACACCUAAACCAGUUCCUCGAACUG-5′(SEQ ID NO: 259)  AAT-864 Target: 5′-ATTGTGGATTTGGTCAAGGAGCTTGAC-3′(SEQ ID NO: 457)  5′-GUGGAUUUGGUCAAGGAGCUUGACA-3′(SEQ ID NO: 656)  3′-AACACCUAAACCAGUUCCUCGAACUGU-5′(SEQ ID NO: 260)  AAT-865 Target: 5′-TTGTGGATTTGGTCAAGGAGCTTGACA-3′(SEQ ID NO: 458)  5′-UGGAUUUGGUCAAGGAGCUUGACAG-3′(SEQ ID NO: 657)  3′-ACACCUAAACCAGUUCCUCGAACUGUC-5′(SEQ ID NO: 261)  AAT-866 Target: 5′-TGTGGATTTGGTCAAGGAGCTTGACAG-3′(SEQ ID NO: 459)  5′-GGAUUUGGUCAAGGAGCUUGACAGA-3′(SEQ ID NO: 658)  3′-CACCUAAACCAGUUCCUCGAACUGUCU-5′(SEQ ID NO: 262)  AAT-867 Target: 5′-GTGGATTTGGTCAAGGAGCTTGACAGA-3′(SEQ ID NO: 460)  5′-GAUUUGGUCAAGGAGCUUGACAGAG-3′(SEQ ID NO: 659)  3′-ACCUAAACCAGUUCCUCGAACUGUCUC-5′(SEQ ID NO: 263)  AAT-868 Target: 5′-TGGATTTGGTCAAGGAGCTTGACAGAG-3′(SEQ ID NO: 461)  5′-AUUUGGUCAAGGAGCUUGACAGAGA-3′(SEQ ID NO: 660)  3′-CCUAAACCAGUUCCUCGAACUGUCUCU-5′(SEQ ID NO: 264)  AAT-869 Target: 5′-GGATTTGGTCAAGGAGCTTGACAGAGA-3′(SEQ ID NO: 462)  5′-UUUGGUCAAGGAGCUUGACAGAGAC-3′(SEQ ID NO: 661)  3′-CUAAACCAGUUCCUCGAACUGUCUCUG-5′(SEQ ID NO: 265)  AAT-870 Target: 5′-GATTTGGTCAAGGAGCTTGACAGAGAC-3′(SEQ ID NO: 463)  5′-UUGGUCAAGGAGCUUGACAGAGACA-3′(SEQ ID NO: 662)  3′-UAAACCAGUUCCUCGAACUGUCUCUGU-5′(SEQ ID NO: 266)  AAT-871 Target: 5′-ATTTGGTCAAGGAGCTTGACAGAGACA-3′(SEQ ID NO: 464)  5′-UGGUCAAGGAGCUUGACAGAGACAC-3′(SEQ ID NO: 663)  3′-AAACCAGUUCCUCGAACUGUCUCUGUG-5′(SEQ ID NO: 267)  AAT-872 Target: 5′-TTTGGTCAAGGAGCTTGACAGAGACAC-3′(SEQ ID NO: 465)  5′-CAGUUUUUGCUCUGGUGAAUUACAU-3′(SEQ ID NO: 664)  3′-GUGUCAAAAACGAGACCACUUAAUGUA-5′(SEQ ID NO: 268)  AAT-896 Target: 5′-CACAGTTTTTGCTCTGGTGAATTACAT-3′(SEQ ID NO: 466)  5′-AGUUUUUGCUCUGGUGAAUUACAUC-3′(SEQ ID NO: 665)  3′-UGUCAAAAACGAGACCACUUAAUGUAG-5′(SEQ ID NO: 269)  AAT-897 Target: 5′-ACAGTTTTTGCTCTGGTGAATTACATC-3′(SEQ ID NO: 467)  5′-GUUUUUGCUCUGGUGAAUUACAUCU-3′(SEQ ID NO: 666)  3′-GUCAAAAACGAGACCACUUAAUGUAGA-5′(SEQ ID NO: 270)  AAT-898 Target: 5′-CAGTTTTTGCTCTGGTGAATTACATCT-3′(SEQ ID NO: 468)  5′-UUUUUGCUCUGGUGAAUUACAUCUU-3′(SEQ ID NO: 667)  3′-UCAAAAACGAGACCACUUAAUGUAGAA-5′(SEQ ID NO: 271)  AAT-899 Target: 5′-AGTTTTTGCTCTGGTGAATTACATCTT-3′(SEQ ID NO: 469)  5′-AAAGGCAAAUGGGAGAGACCCUUUG-3′(SEQ ID NO: 668)  3′-AAUUUCCGUUUACCCUCUCUGGGAAAC-5′(SEQ ID NO: 272)  AAT-928 Target: 5′-TTAAAGGCAAATGGGAGAGACCCTTTG-3′(SEQ ID NO: 470)  5′-AAGGCAAAUGGGAGAGACCCUUUGA-3′(SEQ ID NO: 669)  3′-AUUUCCGUUUACCCUCUCUGGGAAACU-5′(SEQ ID NO: 273)  AAT-929 Target: 5′-TAAAGGCAAATGGGAGAGACCCTTTGA-3′(SEQ ID NO: 471)  5′-AGGCAAAUGGGAGAGACCCUUUGAA-3′(SEQ ID NO: 670)  3′-UUUCCGUUUACCCUCUCUGGGAAACUU-5′(SEQ ID NO: 274)  AAT-930 Target: 5′-AAAGGCAAATGGGAGAGACCCTTTGAA-3′(SEQ ID NO: 472)  5′-GGCAAAUGGGAGAGACCCUUUGAAG-3′(SEQ ID NO: 671)  3′-UUCCGUUUACCCUCUCUGGGAAACUUC-5′(SEQ ID NO: 275)  AAT-931 Target: 5′-AAGGCAAATGGGAGAGACCCTTTGAAG-3′(SEQ ID NO: 473)  5′-AGGAAGAGGACUUCCACGUGGACCA-3′(SEQ ID NO: 672)  3′-GCUCCUUCUCCUGAAGGUGCACCUGGU-5′(SEQ ID NO: 276)  AAT-968 Target: 5′-CGAGGAAGAGGACTTCCACGTGGACCA-3′(SEQ ID NO: 474)  5′-GGAAGAGGACUUCCACGUGGACCAG-3′(SEQ ID NO: 673)  3′-CUCCUUCUCCUGAAGGUGCACCUGGUC-5′(SEQ ID NO: 277)  AAT-969 Target: 5′-GAGGAAGAGGACTTCCACGTGGACCAG-3′(SEQ ID NO: 475)  5′-GAAGAGGACUUCCACGUGGACCAGG-3′(SEQ ID NO: 674)  3′-UCCUUCUCCUGAAGGUGCACCUGGUCC-5′(SEQ ID NO: 278)  AAT-970 Target: 5′-AGGAAGAGGACTTCCACGTGGACCAGG-3′(SEQ ID NO: 476)  5′-AAGAGGACUUCCACGUGGACCAGGU-3′(SEQ ID NO: 675)  3′-CCUUCUCCUGAAGGUGCACCUGGUCCA-5′(SEQ ID NO: 279)  AAT-971 Target: 5′-GGAAGAGGACTTCCACGTGGACCAGGT-3′(SEQ ID NO: 477)  5′-GAGGACUUCCACGUGGACCAGGUGA-3′(SEQ ID NO: 676)  3′-UUCUCCUGAAGGUGCACCUGGUCCACU-5′(SEQ ID NO: 280)  AAT-973 Target: 5′-AAGAGGACTTCCACGTGGACCAGGTGA-3′(SEQ ID NO: 478)  5′-AGGACUUCCACGUGGACCAGGUGAC-3′(SEQ ID NO: 677)  3′-UCUCCUGAAGGUGCACCUGGUCCACUG-5′(SEQ ID NO: 281)  AAT-974 Target: 5′-AGAGGACTTCCACGTGGACCAGGTGAC-3′(SEQ ID NO: 479)  5′-GACUUCCACGUGGACCAGGUGACCA-3′(SEQ ID NO: 678)  3′-UCCUGAAGGUGCACCUGGUCCACUGGU-5′(SEQ ID NO: 282)  AAT-976 Target: 5′-AGGACTTCCACGTGGACCAGGTGACCA-3′(SEQ ID NO: 480)  5′-GUUUAGGCAUGUUUAACAUCCAGCA-3′(SEQ ID NO: 679)  3′-CGCAAAUCCGUACAAAUUGUAGGUCGU-5′(SEQ ID NO: 283)  AAT-1025 Target: 5′-GCGTTTAGGCATGTTTAACATCCAGCA-3′(SEQ ID NO: 481)  5′-UUUAGGCAUGUUUAACAUCCAGCAC-3′(SEQ ID NO: 680)  3′-GCAAAUCCGUACAAAUUGUAGGUCGUG-5′(SEQ ID NO: 284)  AAT-1026 Target: 5′-CGTTTAGGCATGTTTAACATCCAGCAC-3′(SEQ ID NO: 482)  5′-GCUGUCCAGCUGGGUGCUGCUGAUG-3′(SEQ ID NO: 681)  3′-UUCGACAGGUCGACCCACGACGACUAC-5′(SEQ ID NO: 285)  AAT-1059 Target: 5′-AAGCTGTCCAGCTGGGTGCTGCTGATG-3′(SEQ ID NO: 483)  5′-CUGUCCAGCUGGGUGCUGCUGAUGA-3′(SEQ ID NO: 682)  3′-UCGACAGGUCGACCCACGACGACUACU-5′(SEQ ID NO: 286)  AAT-1060 Target: 5′-AGCTGTCCAGCTGGGTGCTGCTGATGA-3′(SEQ ID NO: 484)  5′-CAAUGCCACCGCCAUCUUCUUCCUG-3′(SEQ ID NO: 683)  3′-CCGUUACGGUGGCGGUAGAAGAAGGAC-5′(SEQ ID NO: 287)  AAT-1095 Target: 5′-GGCAATGCCACCGCCATCTTCTTCCTG-3′(SEQ ID NO: 485)  5′-AAUGCCACCGCCAUCUUCUUCCUGC-3′(SEQ ID NO: 684)  3′-CGUUACGGUGGCGGUAGAAGAAGGACG-5′(SEQ ID NO: 288)  AAT-1096 Target: 5′-GCAATGCCACCGCCATCTTCTTCCTGC-3′(SEQ ID NO: 486)  5′-CCACCGCCAUCUUCUUCCUGCCUGA-3′(SEQ ID NO: 685)  3′-ACGGUGGCGGUAGAAGAAGGACGGACU-5′(SEQ ID NO: 289)  AAT-1100 Target: 5′-TGCCACCGCCATCTTCTTCCTGCCTGA-3′(SEQ ID NO: 487)  5′-CACCGCCAUCUUCUUCCUGCCUGAU-3′(SEQ ID NO: 686)  3′-CGGUGGCGGUAGAAGAAGGACGGACUA-5′(SEQ ID NO: 290)  AAT-1101 Target: 5′-GCCACCGCCATCTTCTTCCTGCCTGAT-3′(SEQ ID NO: 488)  5′-ACCGCCAUCUUCUUCCUGCCUGAUG-3′(SEQ ID NO: 687)  3′-GGUGGCGGUAGAAGAAGGACGGACUAC-5′(SEQ ID NO: 291)  AAT-1102 Target: 5′-CCACCGCCATCTTCTTCCTGCCTGATG-3′(SEQ ID NO: 489)  5′-CCGCCAUCUUCUUCCUGCCUGAUGA-3′(SEQ ID NO: 688)  3′-GUGGCGGUAGAAGAAGGACGGACUACU-5′(SEQ ID NO: 292)  AAT-1103 Target: 5′-CACCGCCATCTTCTTCCTGCCTGATGA-3′(SEQ ID NO: 490)  5′-CGCCAUCUUCUUCCUGCCUGAUGAG-3′(SEQ ID NO: 689)  3′-UGGCGGUAGAAGAAGGACGGACUACUC-5′(SEQ ID NO: 293)  AAT-1104 Target: 5′-ACCGCCATCTTCTTCCTGCCTGATGAG-3′(SEQ ID NO: 491)  5′-GCCAUCUUCUUCCUGCCUGAUGAGG-3′(SEQ ID NO: 690)  3′-GGCGGUAGAAGAAGGACGGACUACUCC-5′(SEQ ID NO: 294)  AAT-1105 Target: 5′-CCGCCATCTTCTTCCTGCCTGATGAGG-3′(SEQ ID NO: 492)  5′-AUCUUCUUCCUGCCUGAUGAGGGGA-3′(SEQ ID NO: 691)  3′-GGUAGAAGAAGGACGGACUACUCCCCU-5′(SEQ ID NO: 295)  AAT-1108 Target: 5′-CCATCTTCTTCCTGCCTGATGAGGGGA-3′(SEQ ID NO: 493)  5′-CUUCCUGCCUGAUGAGGGGAAACUA-3′(SEQ ID NO: 692)  3′-AAGAAGGACGGACUACUCCCCUUUGAU-5′(SEQ ID NO: 296)  AAT-1113 Target: 5′-TTCTTCCTGCCTGATGAGGGGAAACTA-3′(SEQ ID NO: 494)  5′-UUCCUGCCUGAUGAGGGGAAACUAC-3′(SEQ ID NO: 693)  3′-AGAAGGACGGACUACUCCCCUUUGAUG-5′(SEQ ID NO: 297)  AAT-1114 Target: 5′-TCTTCCTGCCTGATGAGGGGAAACTAC-3′(SEQ ID NO: 495)  5′-UCCUGCCUGAUGAGGGGAAACUACA-3′(SEQ ID NO: 694)  3′-GAAGGACGGACUACUCCCCUUUGAUGU-5′(SEQ ID NO: 298)  AAT-1115 Target: 5′-CTTCCTGCCTGATGAGGGGAAACTACA-3′(SEQ ID NO: 496)  5′-CCUGCCUGAUGAGGGGAAACUACAG-3′(SEQ ID NO: 695)  3′-AAGGACGGACUACUCCCCUUUGAUGUC-5′(SEQ ID NO: 299)  AAT-1116 Target: 5′-TTCCTGCCTGATGAGGGGAAACTACAG-3′(SEQ ID NO: 497)  5′-CUGCCUGAUGAGGGGAAACUACAGC-3′(SEQ ID NO: 696)  3′-AGGACGGACUACUCCCCUUUGAUGUCG-5′(SEQ ID NO: 300)  AAT-1117 Target: 5′-TCCTGCCTGATGAGGGGAAACTACAGC-3′(SEQ ID NO: 498)  5′-UGCCUGAUGAGGGGAAACUACAGCA-3′(SEQ ID NO: 697)  3′-GGACGGACUACUCCCCUUUGAUGUCGU-5′(SEQ ID NO: 301)  AAT-1118 Target: 5′-CCTGCCTGATGAGGGGAAACTACAGCA-3′(SEQ ID NO: 499)  5′-AGCACCUGGAAAAUGAACUCACCCA-3′(SEQ ID NO: 698)  3′-UGUCGUGGACCUUUUACUUGAGUGGGU-5′(SEQ ID NO: 302)  AAT-1139 Target: 5′-ACAGCACCTGGAAAATGAACTCACCCA-3′(SEQ ID NO: 500)  5′-GCACCUGGAAAAUGAACUCACCCAC-3′(SEQ ID NO: 699)  3′-GUCGUGGACCUUUUACUUGAGUGGGUG-5′(SEQ ID NO: 303)  AAT-1140 Target: 5′-CAGCACCTGGAAAATGAACTCACCCAC-3′(SEQ ID NO: 501)  5′-CACCUGGAAAAUGAACUCACCCACG-3′(SEQ ID NO: 700)  3′-UCGUGGACCUUUUACUUGAGUGGGUGC-5′(SEQ ID NO: 304)  AAT-1141 Target: 5′-AGCACCTGGAAAATGAACTCACCCACG-3′(SEQ ID NO: 502)  5′-ACCUGGAAAAUGAACUCACCCACGA-3′(SEQ ID NO: 701)  3′-CGUGGACCUUUUACUUGAGUGGGUGCU-5′(SEQ ID NO: 305)  AAT-1142 Target: 5′-GCACCTGGAAAATGAACTCACCCACGA-3′(SEQ ID NO: 503)  5′-CCUGGAAAAUGAACUCACCCACGAU-3′(SEQ ID NO: 702)  3′-GUGGACCUUUUACUUGAGUGGGUGCUA-5′(SEQ ID NO: 306)  AAT-1143 Target: 5′-CACCTGGAAAATGAACTCACCCACGAT-3′(SEQ ID NO: 504)  5′-AUAUCAUCACCAAGUUCCUGGAAAA-3′(SEQ ID NO: 703)  3′-GCUAUAGUAGUGGUUCAAGGACCUUUU-5′(SEQ ID NO: 307)  AAT-1166 Target: 5′-CGATATCATCACCAAGTTCCTGGAAAA-3′(SEQ ID NO: 505)  5′-UAUCAUCACCAAGUUCCUGGAAAAU-3′(SEQ ID NO: 704)  3′-CUAUAGUAGUGGUUCAAGGACCUUUUA-5′(SEQ ID NO: 308)  AAT-1167 Target: 5′-GATATCATCACCAAGTTCCTGGAAAAT-3′(SEQ ID NO: 506)  5′-AUCAUCACCAAGUUCCUGGAAAAUG-3′(SEQ ID NO: 705)  3′-UAUAGUAGUGGUUCAAGGACCUUUUAC-5′(SEQ ID NO: 309)  AAT-1168 Target: 5′-ATATCATCACCAAGTTCCTGGAAAATG-3′(SEQ ID NO: 507)  5′-UCAUCACCAAGUUCCUGGAAAAUGA-3′(SEQ ID NO: 706)  3′-AUAGUAGUGGUUCAAGGACCUUUUACU-5′(SEQ ID NO: 310)  AAT-1169 Target: 5′-TATCATCACCAAGTTCCTGGAAAATGA-3′(SEQ ID NO: 508)  5′-CAUCACCAAGUUCCUGGAAAAUGAA-3′(SEQ ID NO: 707)  3′-UAGUAGUGGUUCAAGGACCUUUUACUU-5′(SEQ ID NO: 311)  AAT-1170 Target: 5′-ATCATCACCAAGTTCCTGGAAAATGAA-3′(SEQ ID NO: 509)  5′-AUCACCAAGUUCCUGGAAAAUGAAG-3′(SEQ ID NO: 708)  3′-AGUAGUGGUUCAAGGACCUUUUACUUC-5′(SEQ ID NO: 312)  AAT-1171 Target: 5′-TCATCACCAAGTTCCTGGAAAATGAAG-3′(SEQ ID NO: 510)  5′-UCACCAAGUUCCUGGAAAAUGAAGA-3′(SEQ ID NO: 709)  3′-GUAGUGGUUCAAGGACCUUUUACUUCU-5′(SEQ ID NO: 313)  AAT-1172 Target: 5′-CATCACCAAGTTCCTGGAAAATGAAGA-3′(SEQ ID NO: 511)  5′-CACCAAGUUCCUGGAAAAUGAAGAC-3′(SEQ ID NO: 710)  3′-UAGUGGUUCAAGGACCUUUUACUUCUG-5′(SEQ ID NO: 314)  AAT-1173 Target: 5′-ATCACCAAGTTCCTGGAAAATGAAGAC-3′(SEQ ID NO: 512)  5′-ACCAAGUUCCUGGAAAAUGAAGACA-3′(SEQ ID NO: 711)  3′-AGUGGUUCAAGGACCUUUUACUUCUGU-5′(SEQ ID NO: 315)  AAT-1174 Target: 5′-TCACCAAGTTCCTGGAAAATGAAGACA-3′(SEQ ID NO: 513)  5′-CCAAGUUCCUGGAAAAUGAAGACAG-3′(SEQ ID NO: 712)  3′-GUGGUUCAAGGACCUUUUACUUCUGUC-5′(SEQ ID NO: 316)  AAT-1175 Target: 5′-CACCAAGTTCCTGGAAAATGAAGACAG-3′(SEQ ID NO: 514)  5′-AGGUCUUCAGCAAUGGGGCUGACCU-3′(SEQ ID NO: 713)  3′-AUUCCAGAAGUCGUUACCCCGACUGGA-5′(SEQ ID NO: 317)  AAT-1286 Target: 5′-TAAGGTCTTCAGCAATGGGGCTGACCT-3′(SEQ ID NO: 515)  5′-CAAUGGGGCUGACCUCUCCGGGGUC-3′(SEQ ID NO: 714)  3′-UCGUUACCCCGACUGGAGAGGCCCCAG-5′(SEQ ID NO: 318)  AAT-1296 Target: 5′-AGCAATGGGGCTGACCTCTCCGGGGTC-3′(SEQ ID NO: 516)  5′-AAUGGGGCUGACCUCUCCGGGGUCA-3′(SEQ ID NO: 715)  3′-CGUUACCCCGACUGGAGAGGCCCCAGU-5′(SEQ ID NO: 319)  AAT-1297 Target: 5′-GCAATGGGGCTGACCTCTCCGGGGTCA-3′(SEQ ID NO: 517)  5′-AUGGGGCUGACCUCUCCGGGGUCAC-3′(SEQ ID NO: 716)  3′-GUUACCCCGACUGGAGAGGCCCCAGUG-5′(SEQ ID NO: 320)  AAT-1298 Target: 5′-CAATGGGGCTGACCTCTCCGGGGTCAC-3′(SEQ ID NO: 518)  5′-GAGGAGGCACCCCUGAAGCUCUCCA-3′(SEQ ID NO: 717)  3′-GUCUCCUCCGUGGGGACUUCGAGAGGU-5′(SEQ ID NO: 321)  AAT-1324 Target: 5′-CAGAGGAGGCACCCCTGAAGCTCTCCA-3′(SEQ ID NO: 519)  5′-GGAGGCACCCCUGAAGCUCUCCAAG-3′(SEQ ID NO: 718)  3′-CUCCUCCGUGGGGACUUCGAGAGGUUC-5′(SEQ ID NO: 322)  AAT-1326 Target: 5′-GAGGAGGCACCCCTGAAGCTCTCCAAG-3′(SEQ ID NO: 520)  5′-CUGAAGCUCUCCAAGGCCGUGCAUA-3′(SEQ ID NO: 719)  3′-GGGACUUCGAGAGGUUCCGGCACGUAU-5′(SEQ ID NO: 323)  AAT-1336 Target: 5′-CCCTGAAGCTCTCCAAGGCCGTGCATA-3′(SEQ ID NO: 521)  5′-CGUGCAUAAGGCUGUGCUGACCAUC-3′(SEQ ID NO: 720)  3′-CGGCACGUAUUCCGACACGACUGGUAG-5′(SEQ ID NO: 324)  AAT-1353 Target: 5′-GCCGTGCATAAGGCTGTGCTGACCATC-3′(SEQ ID NO: 522)  5′-GUGCAUAAGGCUGUGCUGACCAUCG-3′(SEQ ID NO: 721)  3′-GGCACGUAUUCCGACACGACUGGUAGC-5′(SEQ ID NO: 325)  AAT-1354 Target: 5′-CCGTGCATAAGGCTGTGCTGACCATCG-3′(SEQ ID NO: 523)  5′-UGCAUAAGGCUGUGCUGACCAUCGA-3′(SEQ ID NO: 722)  3′-GCACGUAUUCCGACACGACUGGUAGCU-5′(SEQ ID NO: 326)  AAT-1355 Target: 5′-CGTGCATAAGGCTGTGCTGACCATCGA-3′(SEQ ID NO: 524)  5′-GCAUAAGGCUGUGCUGACCAUCGAC-3′(SEQ ID NO: 723)  3′-CACGUAUUCCGACACGACUGGUAGCUG-5′(SEQ ID NO: 327)  AAT-1356 Target: 5′-GTGCATAAGGCTGTGCTGACCATCGAC-3′(SEQ ID NO: 525)  5′-CAUAAGGCUGUGCUGACCAUCGACG-3′(SEQ ID NO: 724)  3′-ACGUAUUCCGACACGACUGGUAGCUGC-5′(SEQ ID NO: 328)  AAT-1357 Target: 5′-TGCATAAGGCTGTGCTGACCATCGACG-3′(SEQ ID NO: 526)  5′-AUAAGGCUGUGCUGACCAUCGACGA-3′(SEQ ID NO: 725)  3′-CGUAUUCCGACACGACUGGUAGCUGCU-5′(SEQ ID NO: 329)  AAT-1358 Target: 5′-GCATAAGGCTGTGCTGACCATCGACGA-3′(SEQ ID NO: 527)  5′-UAAGGCUGUGCUGACCAUCGACGAG-3′(SEQ ID NO: 726)  3′-GUAUUCCGACACGACUGGUAGCUGCUC-5′(SEQ ID NO: 330)  AAT-1359 Target: 5′-CATAAGGCTGTGCTGACCATCGACGAG-3′(SEQ ID NO: 528)  5′-AAGGCUGUGCUGACCAUCGACGAGA-3′(SEQ ID NO: 727)  3′-UAUUCCGACACGACUGGUAGCUGCUCU-5′(SEQ ID NO: 331)  AAT-1360 Target: 5′-ATAAGGCTGTGCTGACCATCGACGAGA-3′(SEQ ID NO: 529)  5′-AGGCUGUGCUGACCAUCGACGAGAA-3′(SEQ ID NO: 728)  3′-AUUCCGACACGACUGGUAGCUGCUCUU-5′(SEQ ID NO: 332)  AAT-1361 Target: 5′-TAAGGCTGTGCTGACCATCGACGAGAA-3′(SEQ ID NO: 530)  5′-ACUGAAGCUGCUGGGGCCAUGUUUU-3′(SEQ ID NO: 729)  3′-CCUGACUUCGACGACCCCGGUACAAAA-5′(SEQ ID NO: 333)  AAT-1390 Target: 5′-GGACTGAAGCTGCTGGGGCCATGTTTT-3′(SEQ ID NO: 531)  5′-CUGAAGCUGCUGGGGCCAUGUUUUU-3′(SEQ ID NO: 730)  3′-CUGACUUCGACGACCCCGGUACAAAAA-5′(SEQ ID NO: 334)  AAT-1391 Target: 5′-GACTGAAGCTGCTGGGGCCATGTTTTT-3′(SEQ ID NO: 532)  5′-UGAAGCUGCUGGGGCCAUGUUUUUA-3′(SEQ ID NO: 731)  3′-UGACUUCGACGACCCCGGUACAAAAAU-5′(SEQ ID NO: 335)  AAT-1392 Target: 5′-ACTGAAGCTGCTGGGGCCATGTTTTTA-3′(SEQ ID NO: 533)  5′-GAAGCUGCUGGGGCCAUGUUUUUAG-3′(SEQ ID NO: 732)  3′-GACUUCGACGACCCCGGUACAAAAAUC-5′(SEQ ID NO: 336)  AAT-1393 Target: 5′-CTGAAGCTGCTGGGGCCATGTTTTTAG-3′(SEQ ID NO: 534)  5′-AAGCUGCUGGGGCCAUGUUUUUAGA-3′(SEQ ID NO: 733)  3′-ACUUCGACGACCCCGGUACAAAAAUCU-5′(SEQ ID NO: 337)  AAT-1394 Target: 5′-TGAAGCTGCTGGGGCCATGTTTTTAGA-3′(SEQ ID NO: 535)  5′-AGCUGCUGGGGCCAUGUUUUUAGAG-3′(SEQ ID NO: 734)  3′-CUUCGACGACCCCGGUACAAAAAUCUC-5′(SEQ ID NO: 338)  AAT-1395 Target: 5′-GAAGCTGCTGGGGCCATGTTTTTAGAG-3′(SEQ ID NO: 536)  5′-GCCAUGUUUUUAGAGGCCAUACCCA-3′(SEQ ID NO: 735)  3′-CCCGGUACAAAAAUCUCCGGUAUGGGU-5′(SEQ ID NO: 339)  AAT-1405 Target: 5′-GGGCCATGTTTTTAGAGGCCATACCCA-3′(SEQ ID NO: 537)  5′-CCAUGUUUUUAGAGGCCAUACCCAU-3′(SEQ ID NO: 736)  3′-CCGGUACAAAAAUCUCCGGUAUGGGUA-5′(SEQ ID NO: 340)  AAT-1406 Target: 5′-GGCCATGTTTTTAGAGGCCATACCCAT-3′(SEQ ID NO: 538)  5′-CAUGUUUUUAGAGGCCAUACCCAUG-3′(SEQ ID NO: 737)  3′-CGGUACAAAAAUCUCCGGUAUGGGUAC-5′(SEQ ID NO: 341)  AAT-1407 Target: 5′-GCCATGTTTTTAGAGGCCATACCCATG-3′(SEQ ID NO: 539)  5′-AUGUUUUUAGAGGCCAUACCCAUGU-3′(SEQ ID NO: 738)  3′-GGUACAAAAAUCUCCGGUAUGGGUACA-5′(SEQ ID NO: 342)  AAT-1408 Target: 5′-CCATGTTTTTAGAGGCCATACCCATGT-3′(SEQ ID NO: 540)  5′-UGUUUUUAGAGGCCAUACCCAUGUC-3′(SEQ ID NO: 739)  3′-GUACAAAAAUCUCCGGUAUGGGUACAG-5′(SEQ ID NO: 343)  AAT-1409 Target: 5′-CATGTTTTTAGAGGCCATACCCATGTC-3′(SEQ ID NO: 541)  5′-GUUUUUAGAGGCCAUACCCAUGUCU-3′(SEQ ID NO: 740)  3′-UACAAAAAUCUCCGGUAUGGGUACAGA-5′(SEQ ID NO: 344)  AAT-1410 Target: 5′-ATGTTTTTAGAGGCCATACCCATGTCT-3′(SEQ ID NO: 542)  5′-UUUUUAGAGGCCAUACCCAUGUCUA-3′(SEQ ID NO: 741)  3′-ACAAAAAUCUCCGGUAUGGGUACAGAU-5′(SEQ ID NO: 345)  AAT-1411 Target: 5′-TGTTTTTAGAGGCCATACCCATGTCTA-3′(SEQ ID NO: 543)  5′-UUUUAGAGGCCAUACCCAUGUCUAU-3′(SEQ ID NO: 742)  3′-CAAAAAUCUCCGGUAUGGGUACAGAUA-5′(SEQ ID NO: 346)  AAT-1412 Target: 5′-GTTTTTAGAGGCCATACCCATGTCTAT-3′(SEQ ID NO: 544)  5′-UUUAGAGGCCAUACCCAUGUCUAUC-3′(SEQ ID NO: 743)  3′-AAAAAUCUCCGGUAUGGGUACAGAUAG-5′(SEQ ID NO: 347)  AAT-1413 Target: 5′-TTTTTAGAGGCCATACCCATGTCTATC-3′(SEQ ID NO: 545)  5′-UUAGAGGCCAUACCCAUGUCUAUCC-3′(SEQ ID NO: 744)  3′-AAAAUCUCCGGUAUGGGUACAGAUAGG-5′(SEQ ID NO: 348)  AAT-1414 Target: 5′-TTTTAGAGGCCATACCCATGTCTATCC-3′(SEQ ID NO: 546)  5′-UAGAGGCCAUACCCAUGUCUAUCCC-3′(SEQ ID NO: 745)  3′-AAAUCUCCGGUAUGGGUACAGAUAGGG-5′(SEQ ID NO: 349)  AAT-1415 Target: 5′-TTTAGAGGCCATACCCATGTCTATCCC-3′(SEQ ID NO: 547)  5′-AGAGGCCAUACCCAUGUCUAUCCCC-3′(SEQ ID NO: 746)  3′-AAUCUCCGGUAUGGGUACAGAUAGGGG-5′(SEQ ID NO: 350)  AAT-1416 Target: 5′-TTAGAGGCCATACCCATGTCTATCCCC-3′(SEQ ID NO: 548)  5′-GUUCAACAAACCCUUUGUCUUCUUA-3′(SEQ ID NO: 747)  3′-UUCAAGUUGUUUGGGAAACAGAAGAAU-5′(SEQ ID NO: 351)  AAT-1452 Target: 5′-AAGTTCAACAAACCCTTTGTCTTCTTA-3′(SEQ ID NO: 549)  5′-UUCAACAAACCCUUUGUCUUCUUAA-3′(SEQ ID NO: 748)  3′-UCAAGUUGUUUGGGAAACAGAAGAAUU-5′(SEQ ID NO: 352)  AAT-1453 Target: 5′-AGTTCAACAAACCCTTTGTCTTCTTAA-3′(SEQ ID NO: 550)  5′-UCAACAAACCCUUUGUCUUCUUAAU-3′(SEQ ID NO: 749)  3′-CAAGUUGUUUGGGAAACAGAAGAAUUA-5′(SEQ ID NO: 353)  AAT-1454 Target: 5′-GTTCAACAAACCCTTTGTCTTCTTAAT-3′(SEQ ID NO: 551)  5′-CAACAAACCCUUUGUCUUCUUAAUG-3′(SEQ ID NO: 750)  3′-AAGUUGUUUGGGAAACAGAAGAAUUAC-5′(SEQ ID NO: 354)  AAT-1455 Target: 5′-TTCAACAAACCCTTTGTCTTCTTAATG-3′(SEQ ID NO: 552)  5′-AACAAACCCUUUGUCUUCUUAAUGA-3′(SEQ ID NO: 751)  3′-AGUUGUUUGGGAAACAGAAGAAUUACU-5′(SEQ ID NO: 355)  AAT-1456 Target: 5′-TCAACAAACCCTTTGTCTTCTTAATGA-3′(SEQ ID NO: 553)  5′-ACAAACCCUUUGUCUUCUUAAUGAU-3′(SEQ ID NO: 752)  3′-GUUGUUUGGGAAACAGAAGAAUUACUA-5′(SEQ ID NO: 356)  AAT-1457 Target: 5′-CAACAAACCCTTTGTCTTCTTAATGAT-3′(SEQ ID NO: 554)  5′-CAAACCCUUUGUCUUCUUAAUGAUU-3′(SEQ ID NO: 753)  3′-UUGUUUGGGAAACAGAAGAAUUACUAA-5′(SEQ ID NO: 357)  AAT-1458 Target: 5′-AACAAACCCTTTGTCTTCTTAATGATT-3′(SEQ ID NO: 555)  5′-AAACCCUUUGUCUUCUUAAUGAUUG-3′(SEQ ID NO: 754)  3′-UGUUUGGGAAACAGAAGAAUUACUAAC-5′(SEQ ID NO: 358)  AAT-1459 Target: 5′-ACAAACCCTTTGTCTTCTTAATGATTG-3′(SEQ ID NO: 556)  5′-AACCCUUUGUCUUCUUAAUGAUUGA-3′(SEQ ID NO: 755)  3′-GUUUGGGAAACAGAAGAAUUACUAACU-5′(SEQ ID NO: 359)  AAT-1460 Target: 5′-CAAACCCTTTGTCTTCTTAATGATTGA-3′(SEQ ID NO: 557)  5′-AAUACCAAGUCUCCCCUCUUCAUGG-3′(SEQ ID NO: 756)  3′-UUUUAUGGUUCAGAGGGGAGAAGUACC-5′(SEQ ID NO: 360)  AAT-1489 Target: 5′-AAAATACCAAGTCTCCCCTCTTCATGG-3′(SEQ ID NO: 558)  5′-AUACCAAGUCUCCCCUCUUCAUGGG-3′(SEQ ID NO: 757)  3′-UUUAUGGUUCAGAGGGGAGAAGUACCC-5′(SEQ ID NO: 361)  AAT-1490 Target: 5′-AAATACCAAGTCTCCCCTCTTCATGGG-3′(SEQ ID NO: 559)  5′-UACCAAGUCUCCCCUCUUCAUGGGA-3′(SEQ ID NO: 758)  3′-UUAUGGUUCAGAGGGGAGAAGUACCCU-5′(SEQ ID NO: 362)  AAT-1491 Target: 5′-AATACCAAGTCTCCCCTCTTCATGGGA-3′(SEQ ID NO: 560)  5′-ACCAAGUCUCCCCUCUUCAUGGGAA-3′(SEQ ID NO: 759)  3′-UAUGGUUCAGAGGGGAGAAGUACCCUU-5′(SEQ ID NO: 363)  AAT-1492 Target: 5′-ATACCAAGTCTCCCCTCTTCATGGGAA-3′(SEQ ID NO: 561)  5′-CCAAGUCUCCCCUCUUCAUGGGAAA-3′(SEQ ID NO: 760)  3′-AUGGUUCAGAGGGGAGAAGUACCCUUU-5′(SEQ ID NO: 364)  AAT-1493 Target: 5′-TACCAAGTCTCCCCTCTTCATGGGAAA-3′(SEQ ID NO: 562)  5′-CAAGUCUCCCCUCUUCAUGGGAAAA-3′(SEQ ID NO: 761)  3′-UGGUUCAGAGGGGAGAAGUACCCUUUU-5′(SEQ ID NO: 365)  AAT-1494 Target: 5′-ACCAAGTCTCCCCTCTTCATGGGAAAA-3′(SEQ ID NO: 563)  5′-AAGUCUCCCCUCUUCAUGGGAAAAG-3′(SEQ ID NO: 762)  3′-GGUUCAGAGGGGAGAAGUACCCUUUUC-5′(SEQ ID NO: 366)  AAT-1495 Target: 5′-CCAAGTCTCCCCTCTTCATGGGAAAAG-3′(SEQ ID NO: 564)  5′-AGUCUCCCCUCUUCAUGGGAAAAGU-3′(SEQ ID NO: 763)  3′-GUUCAGAGGGGAGAAGUACCCUUUUCA-5′(SEQ ID NO: 367)  AAT-1496 Target: 5′-CAAGTCTCCCCTCTTCATGGGAAAAGT-3′(SEQ ID NO: 565)  5′-GUCUCCCCUCUUCAUGGGAAAAGUG-3′(SEQ ID NO: 764)  3′-UUCAGAGGGGAGAAGUACCCUUUUCAC-5′(SEQ ID NO: 368)  AAT-1497 Target: 5′-AAGTCTCCCCTCTTCATGGGAAAAGTG-3′(SEQ ID NO: 566)  5′-CUCCCCUCUUCAUGGGAAAAGUGGU-3′(SEQ ID NO: 765)  3′-CAGAGGGGAGAAGUACCCUUUUCACCA-5′(SEQ ID NO: 369)  AAT-1499 Target: 5′-GTCTCCCCTCTTCATGGGAAAAGTGGT-3′(SEQ ID NO: 567)  5′-CCCCUCUUCAUGGGAAAAGUGGUGA-3′(SEQ ID NO: 766)  3′-GAGGGGAGAAGUACCCUUUUCACCACU-5′(SEQ ID NO: 370)  AAT-1501 Target: 5′-CTCCCCTCTTCATGGGAAAAGTGGTGA-3′(SEQ ID NO: 568)  5′-CCCUCUUCAUGGGAAAAGUGGUGAA-3′(SEQ ID NO: 767)  3′-AGGGGAGAAGUACCCUUUUCACCACUU-5′(SEQ ID NO: 371)  AAT-1502 Target: 5′-TCCCCTCTTCATGGGAAAAGTGGTGAA-3′(SEQ ID NO: 569)  5′-CCUCUUCAUGGGAAAAGUGGUGAAU-3′(SEQ ID NO: 768)  3′-GGGGAGAAGUACCCUUUUCACCACUUA-5′(SEQ ID NO: 372)  AAT-1503 Target: 5′-CCCCTCTTCATGGGAAAAGTGGTGAAT-3′(SEQ ID NO: 570)  5′-CUCUUCAUGGGAAAAGUGGUGAAUC-3′(SEQ ID NO: 769)  3′-GGGAGAAGUACCCUUUUCACCACUUAG-5′(SEQ ID NO: 373)  AAT-1504 Target: 5′-CCCTCTTCATGGGAAAAGTGGTGAATC-3′(SEQ ID NO: 571)  5′-UCUUCAUGGGAAAAGUGGUGAAUCC-3′(SEQ ID NO: 770)  3′-GGAGAAGUACCCUUUUCACCACUUAGG-5′(SEQ ID NO: 374)  AAT-1505 Target: 5′-CCTCTTCATGGGAAAAGTGGTGAATCC-3′(SEQ ID NO: 572)  5′-CUUCAUGGGAAAAGUGGUGAAUCCC-3′(SEQ ID NO: 771)  3′-GAGAAGUACCCUUUUCACCACUUAGGG-5′(SEQ ID NO: 375)  AAT-1506 Target: 5′-CTCTTCATGGGAAAAGTGGTGAATCCC-3′(SEQ ID NO: 573)  5′-UUCAUGGGAAAAGUGGUGAAUCCCA-3′(SEQ ID NO: 772)  3′-AGAAGUACCCUUUUCACCACUUAGGGU-5′(SEQ ID NO: 376)  AAT-1507 Target: 5′-TCTTCATGGGAAAAGTGGTGAATCCCA-3′(SEQ ID NO: 574)  5′-UCAUGGGAAAAGUGGUGAAUCCCAC-3′(SEQ ID NO: 773)  3′-GAAGUACCCUUUUCACCACUUAGGGUG-5′(SEQ ID NO: 377)  AAT-1508 Target: 5′-CTTCATGGGAAAAGTGGTGAATCCCAC-3′(SEQ ID NO: 575)  5′-CAUGGGAAAAGUGGUGAAUCCCACC-3′(SEQ ID NO: 774)  3′-AAGUACCCUUUUCACCACUUAGGGUGG-5′(SEQ ID NO: 378)  AAT-1509 Target: 5′-TTCATGGGAAAAGTGGTGAATCCCACC-3′(SEQ ID NO: 576)  5′-AUGGGAAAAGUGGUGAAUCCCACCC-3′(SEQ ID NO: 775)  3′-AGUACCCUUUUCACCACUUAGGGUGGG-5′(SEQ ID NO: 379)  AAT-1510 Target: 5′-TCATGGGAAAAGTGGTGAATCCCACCC-3′(SEQ ID NO: 577)  5′-UGGGAAAAGUGGUGAAUCCCACCCA-3′(SEQ ID NO: 776)  3′-GUACCCUUUUCACCACUUAGGGUGGGU-5′(SEQ ID NO: 380)  AAT-1511 Target: 5′-CATGGGAAAAGTGGTGAATCCCACCCA-3′(SEQ ID NO: 578)  5′-GGGAAAAGUGGUGAAUCCCACCCAA-3′(SEQ ID NO: 777)  3′-UACCCUUUUCACCACUUAGGGUGGGUU-5′(SEQ ID NO: 381)  AAT-1512 Target: 5′-ATGGGAAAAGTGGTGAATCCCACCCAA-3′(SEQ ID NO: 579)  5′-GGAAAAGUGGUGAAUCCCACCCAAA-3′(SEQ ID NO: 778)  3′-ACCCUUUUCACCACUUAGGGUGGGUUU-5′(SEQ ID NO: 382)  AAT-1513 Target: 5′-TGGGAAAAGTGGTGAATCCCACCCAAA-3′(SEQ ID NO: 580)  5′-GAAAAGUGGUGAAUCCCACCCAAAA-3′(SEQ ID NO: 779)  3′-CCCUUUUCACCACUUAGGGUGGGUUUU-5′(SEQ ID NO: 383)  AAT-1514 Target: 5′-GGGAAAAGTGGTGAATCCCACCCAAAA-3′(SEQ ID NO: 581)  5′-AAAAGUGGUGAAUCCCACCCAAAAA-3′(SEQ ID NO: 780)  3′-CCUUUUCACCACUUAGGGUGGGUUUUU-5′(SEQ ID NO: 384)  AAT-1515 Target: 5′-GGAAAAGTGGTGAATCCCACCCAAAAA-3′(SEQ ID NO: 582)  5′-AAAGUGGUGAAUCCCACCCAAAAAU-3′(SEQ ID NO: 781)  3′-CUUUUCACCACUUAGGGUGGGUUUUUA-5′(SEQ ID NO: 385)  AAT-1516 Target: 5′-GAAAAGTGGTGAATCCCACCCAAAAAT-3′(SEQ ID NO: 583)  5′-AAGUGGUGAAUCCCACCCAAAAAUA-3′(SEQ ID NO: 782)  3′-UUUUCACCACUUAGGGUGGGUUUUUAU-5′(SEQ ID NO: 386)  AAT-1517 Target: 5′-AAAAGTGGTGAATCCCACCCAAAAATA-3′(SEQ ID NO: 584)  5′-CGAUAGUUCAAAAUGGUGAAAUUAG-3′(SEQ ID NO: 783)  3′-AAGCUAUCAAGUUUUACCACUUUAAUC-5′(SEQ ID NO: 387)  AAT-2872 Target: 5′-TTCGATAGTTCAAAATGGTGAAATTAG-3′(SEQ ID NO: 585)  5′-CAAAAUGGUGAAAUUAGCAAUUCUA-3′(SEQ ID NO: 784)  3′-AAGUUUUACCACUUUAAUCGUUAAGAU-5′(SEQ ID NO: 388)  AAT-2880 Target: 5′-TTCAAAATGGTGAAATTAGCAATTCTA-3′(SEQ ID NO: 586)  5′-UUGGUAUGAUGUUCAAGUUAGAUAA-3′(SEQ ID NO: 785)  3′-UCAACCAUACUACAAGUUCAAUCUAUU-5′(SEQ ID NO: 389)  AAT-3167 Target: 5′-AGTTGGTATGATGTTCAAGTTAGATAA-3′(SEQ ID NO: 587)  5′-GGUAUGAUGUUCAAGUUAGAUAACA-3′(SEQ ID NO: 786)  3′-AACCAUACUACAAGUUCAAUCUAUUGU-5′(SEQ ID NO: 390)  AAT-3169 Target: 5′-TTGGTATGATGTTCAAGTTAGATAACA-3′(SEQ ID NO: 588)  5′-GUAUGAUGUUCAAGUUAGAUAACAA-3′(SEQ ID NO: 787)  3′-ACCAUACUACAAGUUCAAUCUAUUGUU-5′(SEQ ID NO: 391)  AAT-3170 Target: 5′-TGGTATGATGTTCAAGTTAGATAACAA-3′(SEQ ID NO: 589)  5′-AUGAUGUUCAAGUUAGAUAACAAAA-3′(SEQ ID NO: 788)  3′-CAUACUACAAGUUCAAUCUAUUGUUUU-5′(SEQ ID NO: 392)  AAT-3172 Target: 5′-GTATGATGTTCAAGTTAGATAACAAAA-3′(SEQ ID NO: 590)  5′-AUGUUCAAGUUAGAUAACAAAAUGU-3′(SEQ ID NO: 789)  3′-ACUACAAGUUCAAUCUAUUGUUUUACA-5′(SEQ ID NO: 393)  AAT-3175 Target: 5′-TGATGTTCAAGTTAGATAACAAAATGT-3′(SEQ ID NO: 591)  5′-CAAGUUAGAUAACAAAAUGUUUAUA-3′(SEQ ID NO: 790)  3′-AAGUUCAAUCUAUUGUUUUACAAAUAU-5′(SEQ ID NO: 394)  AAT-3180 Target: 5′-TTCAAGTTAGATAACAAAATGTTTATA-3′(SEQ ID NO: 592)  5′-AAGUUAGAUAACAAAAUGUUUAUAC-3′(SEQ ID NO: 791)  3′-AGUUCAAUCUAUUGUUUUACAAAUAUG-5′(SEQ ID NO: 395)  AAT-3181 Target: 5′-TCAAGTTAGATAACAAAATGTTTATAC-3′(SEQ ID NO: 593)  5′-AGUUAGAUAACAAAAUGUUUAUACC-3′(SEQ ID NO: 792)  3′-GUUCAAUCUAUUGUUUUACAAAUAUGG-5′(SEQ ID NO: 396)  AAT-3182 Target: 5′-CAAGTTAGATAACAAAATGTTTATACC-3′(SEQ ID NO: 594)

TABLE 4 DsiRNA Target Sequences (21mers) in α-1 antitrypsin mRNA AAT-395  21 nt Target: 5′-UACAUCCCACCAUGAUCAGGA-3′ (SEQ ID NO: 793) AAT-475  21 nt Target: 5′-GCCAGCUGGCACACCAGUCCA-3′ (SEQ ID NO: 794) AAT-477  21 nt Target: 5′-CAGCUGGCACACCAGUCCAAC-3′ (SEQ ID NO: 795) AAT-480  21 nt Target: 5′-CUGGCACACCAGUCCAACAGC-3′ (SEQ ID NO: 796) AAT-481  21 nt Target: 5′-UGGCACACCAGUCCAACAGCA-3′ (SEQ ID NO: 797) AAT-482  21 nt Target: 5′-GGCACACCAGUCCAACAGCAC-3′ (SEQ ID NO: 798) AAT-483  21 nt Target: 5′-GCACACCAGUCCAACAGCACC-3′ (SEQ ID NO: 799) AAT-484  21 nt Target: 5′-CACACCAGUCCAACAGCACCA-3′ (SEQ ID NO: 800) AAT-500  21 nt Target: 5′-CACCAAUAUCUUCUUCUCCCC-3′ (SEQ ID NO: 801) AAT-501  21 nt Target: 5′-ACCAAUAUCUUCUUCUCCCCA-3′ (SEQ ID NO: 802) AAT-502  21 nt Target: 5′-CCAAUAUCUUCUUCUCCCCAG-3′ (SEQ ID NO: 803) AAT-503  21 nt Target: 5′-CAAUAUCUUCUUCUCCCCAGU-3′ (SEQ ID NO: 804) AAT-504  21 nt Target: 5′-AAUAUCUUCUUCUCCCCAGUG-3′ (SEQ ID NO: 805) AAT-505  21 nt Target: 5′-AUAUCUUCUUCUCCCCAGUGA-3′ (SEQ ID NO: 806) AAT-506  21 nt Target: 5′-UAUCUUCUUCUCCCCAGUGAG-3′ (SEQ ID NO: 807) AAT-507  21 nt Target: 5′-AUCUUCUUCUCCCCAGUGAGC-3′ (SEQ ID NO: 808) AAT-508  21 nt Target: 5′-UCUUCUUCUCCCCAGUGAGCA-3′ (SEQ ID NO: 809) AAT-509  21 nt Target: 5′-CUUCUUCUCCCCAGUGAGCAU-3′ (SEQ ID NO: 810) AAT-510  21 nt Target: 5′-UUCUUCUCCCCAGUGAGCAUC-3′ (SEQ ID NO: 811) AAT-512  21 nt Target: 5′-CUUCUCCCCAGUGAGCAUCGC-3′ (SEQ ID NO: 812) AAT-513  21 nt Target: 5′-UUCUCCCCAGUGAGCAUCGCU-3′ (SEQ ID NO: 813) AAT-515  21 nt Target: 5′-CUCCCCAGUGAGCAUCGCUAC-3′ (SEQ ID NO: 814) AAT-532  21 nt Target: 5′-CUACAGCCUUUGCAAUGCUCU-3′ (SEQ ID NO: 815) AAT-540  21 nt Target: 5′-UUUGCAAUGCUCUCCCUGGGG-3′ (SEQ ID NO: 816) AAT-581  21 nt Target: 5′-UGAAAUCCUGGAGGGCCUGAA-3′ (SEQ ID NO: 817) AAT-582  21 nt Target: 5′-GAAAUCCUGGAGGGCCUGAAU-3′ (SEQ ID NO: 818) AAT-583  21 nt Target: 5′-AAAUCCUGGAGGGCCUGAAUU-3′ (SEQ ID NO: 819) AAT-585  21 nt Target: 5′-AUCCUGGAGGGCCUGAAUUUC-3′ (SEQ ID NO: 820) AAT-586  21 nt Target: 5′-UCCUGGAGGGCCUGAAUUUCA-3′ (SEQ ID NO: 821) AAT-587  21 nt Target: 5′-CCUGGAGGGCCUGAAUUUCAA-3′ (SEQ ID NO: 822) AAT-634  21 nt Target: 5′-UCCAUGAAGGCUUCCAGGAAC-3′ (SEQ ID NO: 823) AAT-637  21 nt Target: 5′-AUGAAGGCUUCCAGGAACUCC-3′ (SEQ ID NO: 824) AAT-638  21 nt Target: 5′-UGAAGGCUUCCAGGAACUCCU-3′ (SEQ ID NO: 825) AAT-671  21 nt Target: 5′-CCAGCCAGACAGCCAGCUCCA-3′ (SEQ ID NO: 826) AAT-673  21 nt Target: 5′-AGCCAGACAGCCAGCUCCAGC-3′ (SEQ ID NO: 827) AAT-675  21 nt Target: 5′-CCAGACAGCCAGCUCCAGCUG-3′ (SEQ ID NO: 828) AAT-676  21 nt Target: 5′-CAGACAGCCAGCUCCAGCUGA-3′ (SEQ ID NO: 829) AAT-734  21 nt Target: 5′-GCUAGUGGAUAAGUUUUUGGA-3′ (SEQ ID NO: 830) AAT-735  21 nt Target: 5′-CUAGUGGAUAAGUUUUUGGAG-3′ (SEQ ID NO: 831) AAT-736  21 nt Target: 5′-UAGUGGAUAAGUUUUUGGAGG-3′ (SEQ ID NO: 832) AAT-737  21 nt Target: 5′-AGUGGAUAAGUUUUUGGAGGA-3′ (SEQ ID NO: 833) AAT-738  21 nt Target: 5′-GUGGAUAAGUUUUUGGAGGAU-3′ (SEQ ID NO: 834) AAT-739  21 nt Target: 5′-UGGAUAAGUUUUUGGAGGAUG-3′ (SEQ ID NO: 835) AAT-740  21 nt Target: 5′-GGAUAAGUUUUUGGAGGAUGU-3′ (SEQ ID NO: 836) AAT-767  21 nt Target: 5′-GUUGUACCACUCAGAAGCCUU-3′ (SEQ ID NO: 837) AAT-768  21 nt Target: 5′-UUGUACCACUCAGAAGCCUUC-3′ (SEQ ID NO: 838) AAT-850  21 nt Target: 5′-GUACUCAAGGGAAAAUUGUGG-3′ (SEQ ID NO: 839) AAT-851  21 nt Target: 5′-UACUCAAGGGAAAAUUGUGGA-3′ (SEQ ID NO: 840) AAT-852  21 nt Target: 5′-ACUCAAGGGAAAAUUGUGGAU-3′ (SEQ ID NO: 841) AAT-853  21 nt Target: 5′-CUCAAGGGAAAAUUGUGGAUU-3′ (SEQ ID NO: 842) AAT-854  21 nt Target: 5′-UCAAGGGAAAAUUGUGGAUUU-3′ (SEQ ID NO: 843) AAT-855  21 nt Target: 5′-CAAGGGAAAAUUGUGGAUUUG-3′ (SEQ ID NO: 844) AAT-856  21 nt Target: 5′-AAGGGAAAAUUGUGGAUUUGG-3′ (SEQ ID NO: 845) AAT-857  21 nt Target: 5′-AGGGAAAAUUGUGGAUUUGGU-3′ (SEQ ID NO: 846) AAT-858  21 nt Target: 5′-GGGAAAAUUGUGGAUUUGGUC-3′ (SEQ ID NO: 847) AAT-859  21 nt Target: 5′-GGAAAAUUGUGGAUUUGGUCA-3′ (SEQ ID NO: 848) AAT-860  21 nt Target: 5′-GAAAAUUGUGGAUUUGGUCAA-3′ (SEQ ID NO: 849) AAT-861  21 nt Target: 5′-AAAAUUGUGGAUUUGGUCAAG-3′ (SEQ ID NO: 850) AAT-862  21 nt Target: 5′-AAAUUGUGGAUUUGGUCAAGG-3′ (SEQ ID NO: 851) AAT-863  21 nt Target: 5′-AAUUGUGGAUUUGGUCAAGGA-3′ (SEQ ID NO: 852) AAT-864  21 nt Target: 5′-AUUGUGGAUUUGGUCAAGGAG-3′ (SEQ ID NO: 853) AAT-865  21 nt Target: 5′-UUGUGGAUUUGGUCAAGGAGC-3′ (SEQ ID NO: 854) AAT-866  21 nt Target: 5′-UGUGGAUUUGGUCAAGGAGCU-3′ (SEQ ID NO: 855) AAT-867  21 nt Target: 5′-GUGGAUUUGGUCAAGGAGCUU-3′ (SEQ ID NO: 856) AAT-868  21 nt Target: 5′-UGGAUUUGGUCAAGGAGCUUG-3′ (SEQ ID NO: 857) AAT-869  21 nt Target: 5′-GGAUUUGGUCAAGGAGCUUGA-3′ (SEQ ID NO: 858) AAT-870  21 nt Target: 5′-GAUUUGGUCAAGGAGCUUGAC-3′ (SEQ ID NO: 859) AAT-871  21 nt Target: 5′-AUUUGGUCAAGGAGCUUGACA-3′ (SEQ ID NO: 860) AAT-872  21 nt Target: 5′-UUUGGUCAAGGAGCUUGACAG-3′ (SEQ ID NO: 861) AAT-896  21 nt Target: 5′-CACAGUUUUUGCUCUGGUGAA-3′ (SEQ ID NO: 862) AAT-897  21 nt Target: 5′-ACAGUUUUUGCUCUGGUGAAU-3′ (SEQ ID NO: 863) AAT-898  21 nt Target: 5′-CAGUUUUUGCUCUGGUGAAUU-3′ (SEQ ID NO: 864) AAT-899  21 nt Target: 5′-AGUUUUUGCUCUGGUGAAUUA-3′ (SEQ ID NO: 865) AAT-928  21 nt Target: 5′-UUAAAGGCAAAUGGGAGAGAC-3′ (SEQ ID NO: 866) AAT-929  21 nt Target: 5′-UAAAGGCAAAUGGGAGAGACC-3′ (SEQ ID NO: 867) AAT-930  21 nt Target: 5′-AAAGGCAAAUGGGAGAGACCC-3′ (SEQ ID NO: 868) AAT-931  21 nt Target: 5′-AAGGCAAAUGGGAGAGACCCU-3′ (SEQ ID NO: 869) AAT-968  21 nt Target: 5′-CGAGGAAGAGGACUUCCACGU-3′ (SEQ ID NO: 870) AAT-969  21 nt Target: 5′-GAGGAAGAGGACUUCCACGUG-3′ (SEQ ID NO: 871) AAT-970  21 nt Target: 5′-AGGAAGAGGACUUCCACGUGG-3′ (SEQ ID NO: 872) AAT-971  21 nt Target: 5′-GGAAGAGGACUUCCACGUGGA-3′ (SEQ ID NO: 873) AAT-973  21 nt Target: 5′-AAGAGGACUUCCACGUGGACC-3′ (SEQ ID NO: 874) AAT-974  21 nt Target: 5′-AGAGGACUUCCACGUGGACCA-3′ (SEQ ID NO: 875) AAT-976  21 nt Target: 5′-AGGACUUCCACGUGGACCAGG-3′ (SEQ ID NO: 876) AAT-1025 21 nt Target: 5′-GCGUUUAGGCAUGUUUAACAU-3′ (SEQ ID NO: 877) AAT-1026 21 nt Target: 5′-CGUUUAGGCAUGUUUAACAUC-3′ (SEQ ID NO: 878) AAT-1059 21 nt Target: 5′-AAGCUGUCCAGCUGGGUGCUG-3′ (SEQ ID NO: 879) AAT-1060 21 nt Target: 5′-AGCUGUCCAGCUGGGUGCUGC-3′ (SEQ ID NO: 880) AAT-1095 21 nt Target: 5′-GGCAAUGCCACCGCCAUCUUC-3′ (SEQ ID NO: 881) AAT-1096 21 nt Target: 5′-GCAAUGCCACCGCCAUCUUCU-3′ (SEQ ID NO: 882) AAT-1100 21 nt Target: 5′-UGCCACCGCCAUCUUCUUCCU-3′ (SEQ ID NO: 883) AAT-1101 21 nt Target: 5′-GCCACCGCCAUCUUCUUCCUG-3′ (SEQ ID NO: 884) AAT-1102 21 nt Target: 5′-CCACCGCCAUCUUCUUCCUGC-3′ (SEQ ID NO: 885) AAT-1103 21 nt Target: 5′-CACCGCCAUCUUCUUCCUGCC-3′ (SEQ ID NO: 886) AAT-1104 21 nt Target: 5′-ACCGCCAUCUUCUUCCUGCCU-3′ (SEQ ID NO: 887) AAT-1105 21 nt Target: 5′-CCGCCAUCUUCUUCCUGCCUG-3′ (SEQ ID NO: 888) AAT-1108 21 nt Target: 5′-CCAUCUUCUUCCUGCCUGAUG-3′ (SEQ ID NO: 889) AAT-1113 21 nt Target: 5′-UUCUUCCUGCCUGAUGAGGGG-3′ (SEQ ID NO: 890) AAT-1114 21 nt Target: 5′-UCUUCCUGCCUGAUGAGGGGA-3′ (SEQ ID NO: 891) AAT-1115 21 nt Target: 5′-CUUCCUGCCUGAUGAGGGGAA-3′ (SEQ ID NO: 892) AAT-1116 21 nt Target: 5′-UUCCUGCCUGAUGAGGGGAAA-3′ (SEQ ID NO: 893) AAT-1117 21 nt Target: 5′-UCCUGCCUGAUGAGGGGAAAC-3′ (SEQ ID NO: 894) AAT-1118 21 nt Target: 5′-CCUGCCUGAUGAGGGGAAACU-3′ (SEQ ID NO: 895) AAT-1139 21 nt Target: 5′-ACAGCACCUGGAAAAUGAACU-3′ (SEQ ID NO: 896) AAT-1140 21 nt Target: 5′-CAGCACCUGGAAAAUGAACUC-3′ (SEQ ID NO: 897) AAT-1141 21 nt Target: 5′-AGCACCUGGAAAAUGAACUCA-3′ (SEQ ID NO: 898) AAT-1142 21 nt Target: 5′-GCACCUGGAAAAUGAACUCAC-3′ (SEQ ID NO: 899) AAT-1143 21 nt Target: 5′-CACCUGGAAAAUGAACUCACC-3′ (SEQ ID NO: 900) AAT-1166 21 nt Target: 5′-CGAUAUCAUCACCAAGUUCCU-3′ (SEQ ID NO: 901) AAT-1167 21 nt Target: 5′-GAUAUCAUCACCAAGUUCCUG-3′ (SEQ ID NO: 902) AAT-1168 21 nt Target: 5′-AUAUCAUCACCAAGUUCCUGG-3′ (SEQ ID NO: 903) AAT-1169 21 nt Target: 5′-UAUCAUCACCAAGUUCCUGGA-3′ (SEQ ID NO: 904) AAT-1170 21 nt Target: 5′-AUCAUCACCAAGUUCCUGGAA-3′ (SEQ ID NO: 905) AAT-1171 21 nt Target: 5′-UCAUCACCAAGUUCCUGGAAA-3′ (SEQ ID NO: 906) AAT-1172 21 nt Target: 5′-CAUCACCAAGUUCCUGGAAAA-3′ (SEQ ID NO: 907) AAT-1173 21 nt Target: 5′-AUCACCAAGUUCCUGGAAAAU-3′ (SEQ ID NO: 908) AAT-1174 21 nt Target: 5′-UCACCAAGUUCCUGGAAAAUG-3′ (SEQ ID NO: 909) AAT-1175 21 nt Target: 5′-CACCAAGUUCCUGGAAAAUGA-3′ (SEQ ID NO: 910) AAT-1286 21 nt Target: 5′-UAAGGUCUUCAGCAAUGGGGC-3′ (SEQ ID NO: 911) AAT-1296 21 nt Target: 5′-AGCAAUGGGGCUGACCUCUCC-3′ (SEQ ID NO: 912) AAT-1297 21 nt Target: 5′-GCAAUGGGGCUGACCUCUCCG-3′ (SEQ ID NO: 913) AAT-1298 21 nt Target: 5′-CAAUGGGGCUGACCUCUCCGG-3′ (SEQ ID NO: 914) AAT-1324 21 nt Target: 5′-CAGAGGAGGCACCCCUGAAGC-3′ (SEQ ID NO: 915) AAT-1326 21 nt Target: 5′-GAGGAGGCACCCCUGAAGCUC-3′ (SEQ ID NO: 916) AAT-1336 21 nt Target: 5′-CCCUGAAGCUCUCCAAGGCCG-3′ (SEQ ID NO: 917) AAT-1353 21 nt Target: 5′-GCCGUGCAUAAGGCUGUGCUG-3′ (SEQ ID NO: 918) AAT-1354 21 nt Target: 5′-CCGUGCAUAAGGCUGUGCUGA-3′ (SEQ ID NO: 919) AAT-1355 21 nt Target: 5′-CGUGCAUAAGGCUGUGCUGAC-3′ (SEQ ID NO: 920) AAT-1356 21 nt Target: 5′-GUGCAUAAGGCUGUGCUGACC-3′ (SEQ ID NO: 921) AAT-1357 21 nt Target: 5′-UGCAUAAGGCUGUGCUGACCA-3′ (SEQ ID NO: 922) AAT-1358 21 nt Target: 5′-GCAUAAGGCUGUGCUGACCAU-3′ (SEQ ID NO: 923) AAT-1359 21 nt Target: 5′-CAUAAGGCUGUGCUGACCAUC-3′ (SEQ ID NO: 924) AAT-1360 21 nt Target: 5′-AUAAGGCUGUGCUGACCAUCG-3′ (SEQ ID NO: 925) AAT-1361 21 nt Target: 5′-UAAGGCUGUGCUGACCAUCGA-3′ (SEQ ID NO: 926) AAT-1390 21 nt Target: 5′-GGACUGAAGCUGCUGGGGCCA-3′ (SEQ ID NO: 927) AAT-1391 21 nt Target: 5′-GACUGAAGCUGCUGGGGCCAU-3′ (SEQ ID NO: 928) AAT-1392 21 nt Target: 5′-ACUGAAGCUGCUGGGGCCAUG-3′ (SEQ ID NO: 929) AAT-1393 21 nt Target: 5′-CUGAAGCUGCUGGGGCCAUGU-3′ (SEQ ID NO: 930) AAT-1394 21 nt Target: 5′-UGAAGCUGCUGGGGCCAUGUU-3′ (SEQ ID NO: 931) AAT-1395 21 nt Target: 5′-GAAGCUGCUGGGGCCAUGUUU-3′ (SEQ ID NO: 932) AAT-1405 21 nt Target: 5′-GGGCCAUGUUUUUAGAGGCCA-3′ (SEQ ID NO: 933) AAT-1406 21 nt Target: 5′-GGCCAUGUUUUUAGAGGCCAU-3′ (SEQ ID NO: 934) AAT-1407 21 nt Target: 5′-GCCAUGUUUUUAGAGGCCAUA-3′ (SEQ ID NO: 935) AAT-1408 21 nt Target: 5′-CCAUGUUUUUAGAGGCCAUAC-3′ (SEQ ID NO: 936) AAT-1409 21 nt Target: 5′-CAUGUUUUUAGAGGCCAUACC-3′ (SEQ ID NO: 937) AAT-1410 21 nt Target: 5′-AUGUUUUUAGAGGCCAUACCC-3′ (SEQ ID NO: 938) AAT-1411 21 nt Target: 5′-UGUUUUUAGAGGCCAUACCCA-3′ (SEQ ID NO: 939) AAT-1412 21 nt Target: 5′-GUUUUUAGAGGCCAUACCCAU-3′ (SEQ ID NO: 940) AAT-1413 21 nt Target: 5′-UUUUUAGAGGCCAUACCCAUG-3′ (SEQ ID NO: 941) AAT-1414 21 nt Target: 5′-UUUUAGAGGCCAUACCCAUGU-3′ (SEQ ID NO: 942) AAT-1415 21 nt Target: 5′-UUUAGAGGCCAUACCCAUGUC-3′ (SEQ ID NO: 943) AAT-1416 21 nt Target: 5′-UUAGAGGCCAUACCCAUGUCU-3′ (SEQ ID NO: 944) AAT-1452 21 nt Target: 5′-AAGUUCAACAAACCCUUUGUC-3′ (SEQ ID NO: 945) AAT-1453 21 nt Target: 5′-AGUUCAACAAACCCUUUGUCU-3′ (SEQ ID NO: 946) AAT-1454 21 nt Target: 5′-GUUCAACAAACCCUUUGUCUU-3′ (SEQ ID NO: 947) AAT-1455 21 nt Target: 5′-UUCAACAAACCCUUUGUCUUC-3′ (SEQ ID NO: 948) AAT-1456 21 nt Target: 5′-UCAACAAACCCUUUGUCUUCU-3′ (SEQ ID NO: 949) AAT-1457 21 nt Target: 5′-CAACAAACCCUUUGUCUUCUU-3′ (SEQ ID NO: 950) AAT-1458 21 nt Target: 5′-AACAAACCCUUUGUCUUCUUA-3′ (SEQ ID NO: 951) AAT-1459 21 nt Target: 5′-ACAAACCCUUUGUCUUCUUAA-3′ (SEQ ID NO: 952) AAT-1460 21 nt Target: 5′-CAAACCCUUUGUCUUCUUAAU-3′ (SEQ ID NO: 953) AAT-1489 21 nt Target: 5′-AAAAUACCAAGUCUCCCCUCU-3′ (SEQ ID NO: 954) AAT-1490 21 nt Target: 5′-AAAUACCAAGUCUCCCCUCUU-3′ (SEQ ID NO: 955) AAT-1491 21 nt Target: 5′-AAUACCAAGUCUCCCCUCUUC-3′ (SEQ ID NO: 956) AAT-1492 21 nt Target: 5′-AUACCAAGUCUCCCCUCUUCA-3′ (SEQ ID NO: 957) AAT-1493 21 nt Target: 5′-UACCAAGUCUCCCCUCUUCAU-3′ (SEQ ID NO: 958) AAT-1494 21 nt Target: 5′-ACCAAGUCUCCCCUCUUCAUG-3′ (SEQ ID NO: 959) AAT-1495 21 nt Target: 5′-CCAAGUCUCCCCUCUUCAUGG-3′ (SEQ ID NO: 960) AAT-1496 21 nt Target: 5′-CAAGUCUCCCCUCUUCAUGGG-3′ (SEQ ID NO: 961) AAT-1497 21 nt Target: 5′-AAGUCUCCCCUCUUCAUGGGA-3′ (SEQ ID NO: 962) AAT-1499 21 nt Target: 5′-GUCUCCCCUCUUCAUGGGAAA-3′ (SEQ ID NO: 963) AAT-1501 21 nt Target: 5′-CUCCCCUCUUCAUGGGAAAAG-3′ (SEQ ID NO: 964) AAT-1502 21 nt Target: 5′-UCCCCUCUUCAUGGGAAAAGU-3′ (SEQ ID NO: 965) AAT-1503 21 nt Target: 5′-CCCCUCUUCAUGGGAAAAGUG-3′ (SEQ ID NO: 966) AAT-1504 21 nt Target: 5′-CCCUCUUCAUGGGAAAAGUGG-3′ (SEQ ID NO: 967) AAT-1505 21 nt Target: 5′-CCUCUUCAUGGGAAAAGUGGU-3′ (SEQ ID NO: 968) AAT-1506 21 nt Target: 5′-CUCUUCAUGGGAAAAGUGGUG-3′ (SEQ ID NO: 969) AAT-1507 21 nt Target: 5′-UCUUCAUGGGAAAAGUGGUGA-3′ (SEQ ID NO: 970) AAT-1508 21 nt Target: 5′-CUUCAUGGGAAAAGUGGUGAA-3′ (SEQ ID NO: 971) AAT-1509 21 nt Target: 5′-UUCAUGGGAAAAGUGGUGAAU-3′ (SEQ ID NO: 972) AAT-1510 21 nt Target: 5′-UCAUGGGAAAAGUGGUGAAUC-3′ (SEQ ID NO: 973) AAT-1511 21 nt Target: 5′-CAUGGGAAAAGUGGUGAAUCC-3′ (SEQ ID NO: 974) AAT-1512 21 nt Target: 5′-AUGGGAAAAGUGGUGAAUCCC-3′ (SEQ ID NO: 975) AAT-1513 21 nt Target: 5′-UGGGAAAAGUGGUGAAUCCCA-3′ (SEQ ID NO: 976) AAT-1514 21 nt Target: 5′-GGGAAAAGUGGUGAAUCCCAC-3′ (SEQ ID NO: 977) AAT-1515 21 nt Target: 5′-GGAAAAGUGGUGAAUCCCACC-3′ (SEQ ID NO: 978) AAT-1516 21 nt Target: 5′-GAAAAGUGGUGAAUCCCACCC-3′ (SEQ ID NO: 979) AAT-1517 21 nt Target: 5′-AAAAGUGGUGAAUCCCACCCA-3′ (SEQ ID NO: 980) AAT-2872 21 nt Target: 5′-UUCGAUAGUUCAAAAUGGUGA-3′ (SEQ ID NO: 981) AAT-2880 21 nt Target: 5′-UUCAAAAUGGUGAAAUUAGCA-3′ (SEQ ID NO: 982) AAT-3167 21 nt Target: 5′-AGUUGGUAUGAUGUUCAAGUU-3′ (SEQ ID NO: 983) AAT-3169 21 nt Target: 5′-UUGGUAUGAUGUUCAAGUUAG-3′ (SEQ ID NO: 984) AAT-3170 21 nt Target: 5′-UGGUAUGAUGUUCAAGUUAGA-3′ (SEQ ID NO: 985) AAT-3172 21 nt Target: 5′-GUAUGAUGUUCAAGUUAGAUA-3′ (SEQ ID NO: 986) AAT-3175 21 nt Target: 5′-UGAUGUUCAAGUUAGAUAACA-3′ (SEQ ID NO: 987) AAT-3180 21 nt Target: 5′-UUCAAGUUAGAUAACAAAAUG-3′ (SEQ ID NO: 988) AAT-3181 21 nt Target: 5′-UCAAGUUAGAUAACAAAAUGU-3′ (SEQ ID NO: 989) AAT-3182 21 nt Target: 5′-CAAGUUAGAUAACAAAAUGUU-3′ (SEQ ID NO: 990)

TABLE 5 Selected Human Anti-α-1 antitrypsin “Blunt/Blunt” DsiRNAs 5′-UACAUCCCACCAUGAUCAGGAUCACCC-3′ (SEQ ID NO: 991) 3′-AUGUAGGGUGGUACUAGUCCUAGUGGG-5′ (SEQ ID NO: 199) AAT-395 Target: 5′-TACATCCCACCATGATCAGGATCACCC-3′ (SEQ ID NO: 397) 5′-GCCAGCUGGCACACCAGUCCAACAGCA-3′ (SEQ ID NO: 992) 3′-CGGUCGACCGUGUGGUCAGGUUGUCGU-5′ (SEQ ID NO: 200) AAT-475 Target: 5′-GCCAGCTGGCACACCAGTCCAACAGCA-3′ (SEQ ID NO: 398) 5′-CAGCUGGCACACCAGUCCAACAGCACC-3′ (SEQ ID NO: 993) 3′-GUCGACCGUGUGGUCAGGUUGUCGUGG-5′ (SEQ ID NO: 201) AAT-477 Target: 5′-CAGCTGGCACACCAGTCCAACAGCACC-3′ (SEQ ID NO: 399) 5′-CUGGCACACCAGUCCAACAGCACCAAU-3′ (SEQ ID NO: 994) 3′-GACCGUGUGGUCAGGUUGUCGUGGUUA-5′ (SEQ ID NO: 202) AAT-480 Target: 5′-CTGGCACACCAGTCCAACAGCACCAAT-3′ (SEQ ID NO: 400) 5′-UGGCACACCAGUCCAACAGCACCAAUA-3′ (SEQ ID NO: 995) 3′-ACCGUGUGGUCAGGUUGUCGUGGUUAU-5′ (SEQ ID NO: 203) AAT-481 Target: 5′-TGGCACACCAGTCCAACAGCACCAATA-3′ (SEQ ID NO: 401) 5′-GGCACACCAGUCCAACAGCACCAAUAU-3′ (SEQ ID NO: 996) 3′-CCGUGUGGUCAGGUUGUCGUGGUUAUA-5′ (SEQ ID NO: 204) AAT-482 Target: 5′-GGCACACCAGTCCAACAGCACCAATAT-3′ (SEQ ID NO: 402) 5′-GCACACCAGUCCAACAGCACCAAUAUC-3′ (SEQ ID NO: 997) 3′-CGUGUGGUCAGGUUGUCGUGGUUAUAG-5′ (SEQ ID NO: 205) AAT-483 Target: 5′-GCACACCAGTCCAACAGCACCAATATC-3′ (SEQ ID NO: 403) 5′-CACACCAGUCCAACAGCACCAAUAUCU-3′ (SEQ ID NO: 998) 3′-GUGUGGUCAGGUUGUCGUGGUUAUAGA-5′ (SEQ ID NO: 206) AAT-484 Target: 5′-CACACCAGTCCAACAGCACCAATATCT-3′ (SEQ ID NO: 404) 5′-CACCAAUAUCUUCUUCUCCCCAGUGAG-3′ (SEQ ID NO: 999) 3′-GUGGUUAUAGAAGAAGAGGGGUCACUC-5′ (SEQ ID NO: 207) AAT-500 Target: 5′-CACCAATATCTTCTTCTCCCCAGTGAG-3′ (SEQ ID NO: 405) 5′-ACCAAUAUCUUCUUCUCCCCAGUGAGC-3′ (SEQ ID NO: 1000) 3′-UGGUUAUAGAAGAAGAGGGGUCACUCG-5′ (SEQ ID NO: 208) AAT-501 Target: 5′-ACCAATATCTTCTTCTCCCCAGTGAGC-3′ (SEQ ID NO: 406) 5′-CCAAUAUCUUCUUCUCCCCAGUGAGCA-3′ (SEQ ID NO: 1001) 3′-GGUUAUAGAAGAAGAGGGGUCACUCGU-5′ (SEQ ID NO: 209) AAT-502 Target: 5′-CCAATATCTTCTTCTCCCCAGTGAGCA-3′ (SEQ ID NO: 407) 5′-CAAUAUCUUCUUCUCCCCAGUGAGCAU-3′ (SEQ ID NO: 1002) 3′-GUUAUAGAAGAAGAGGGGUCACUCGUA-5′ (SEQ ID NO: 210) AAT-503 Target: 5′-CAATATCTTCTTCTCCCCAGTGAGCAT-3′ (SEQ ID NO: 408) 5′-AAUAUCUUCUUCUCCCCAGUGAGCAUC-3′ (SEQ ID NO: 1003) 3′-UUAUAGAAGAAGAGGGGUCACUCGUAG-5′ (SEQ ID NO: 211) AAT-504 Target: 5′-AATATCTTCTTCTCCCCAGTGAGCATC-3′ (SEQ ID NO: 409) 5′-AUAUCUUCUUCUCCCCAGUGAGCAUCG-3′ (SEQ ID NO: 1004) 3′-UAUAGAAGAAGAGGGGUCACUCGUAGC-5′ (SEQ ID NO: 212) AAT-505 Target: 5′-ATATCTTCTTCTCCCCAGTGAGCATCG-3′ (SEQ ID NO: 410) 5′-UAUCUUCUUCUCCCCAGUGAGCAUCGC-3′ (SEQ ID NO: 1005) 3′-AUAGAAGAAGAGGGGUCACUCGUAGCG-5′ (SEQ ID NO: 213) AAT-506 Target: 5′-TATCTTCTTCTCCCCAGTGAGCATCGC-3′ (SEQ ID NO: 411) 5′-AUCUUCUUCUCCCCAGUGAGCAUCGCU-3′ (SEQ ID NO: 1006) 3′-UAGAAGAAGAGGGGUCACUCGUAGCGA-5′ (SEQ ID NO: 214) AAT-507 Target: 5′-ATCTTCTTCTCCCCAGTGAGCATCGCT-3′ (SEQ ID NO: 412) 5′-UCUUCUUCUCCCCAGUGAGCAUCGCUA-3′ (SEQ ID NO: 1007) 3′-AGAAGAAGAGGGGUCACUCGUAGCGAU-5′ (SEQ ID NO: 215) AAT-508 Target: 5′-TCTTCTTCTCCCCAGTGAGCATCGCTA-3′ (SEQ ID NO: 413) 5′-CUUCUUCUCCCCAGUGAGCAUCGCUAC-3′ (SEQ ID NO: 1008) 3′-GAAGAAGAGGGGUCACUCGUAGCGAUG-5′ (SEQ ID NO: 216) AAT-509 Target: 5′-CTTCTTCTCCCCAGTGAGCATCGCTAC-3′ (SEQ ID NO: 414) 5′-UUCUUCUCCCCAGUGAGCAUCGCUACA-3′ (SEQ ID NO: 1009) 3′-AAGAAGAGGGGUCACUCGUAGCGAUGU-5′ (SEQ ID NO: 217) AAT-510 Target: 5′-TTCTTCTCCCCAGTGAGCATCGCTACA-3′ (SEQ ID NO: 415) 5′-CUUCUCCCCAGUGAGCAUCGCUACAGC-3′ (SEQ ID NO: 1010) 3′-GAAGAGGGGUCACUCGUAGCGAUGUCG-5′ (SEQ ID NO: 218) AAT-512 Target: 5′-CTTCTCCCCAGTGAGCATCGCTACAGC-3′ (SEQ ID NO: 416) 5′-UUCUCCCCAGUGAGCAUCGCUACAGCC-3′ (SEQ ID NO: 1011) 3′-AAGAGGGGUCACUCGUAGCGAUGUCGG-5′ (SEQ ID NO: 219) AAT-513 Target: 5′-TTCTCCCCAGTGAGCATCGCTACAGCC-3′ (SEQ ID NO: 417) 5′-CUCCCCAGUGAGCAUCGCUACAGCCUU-3′ (SEQ ID NO: 1012) 3′-GAGGGGUCACUCGUAGCGAUGUCGGAA-5′ (SEQ ID NO: 220) AAT-515 Target: 5′-CTCCCCAGTGAGCATCGCTACAGCCTT-3′ (SEQ ID NO: 418) 5′-CUACAGCCUUUGCAAUGCUCUCCCUGG-3′ (SEQ ID NO: 1013) 3′-GAUGUCGGAAACGUUACGAGAGGGACC-5′ (SEQ ID NO: 221) AAT-532 Target: 5′-CTACAGCCTTTGCAATGCTCTCCCTGG-3′ (SEQ ID NO: 419) 5′-UUUGCAAUGCUCUCCCUGGGGACCAAG-3′ (SEQ ID NO: 1014) 3′-AAACGUUACGAGAGGGACCCCUGGUUC-5′ (SEQ ID NO: 222) AAT-540 Target: 5′-TTTGCAATGCTCTCCCTGGGGACCAAG-3′ (SEQ ID NO: 420) 5′-UGAAAUCCUGGAGGGCCUGAAUUUCAA-3′ (SEQ ID NO: 1015) 3′-ACUUUAGGACCUCCCGGACUUAAAGUU-5′ (SEQ ID NO: 223) AAT-581 Target: 5′-TGAAATCCTGGAGGGCCTGAATTTCAA-3′ (SEQ ID NO: 421) 5′-GAAAUCCUGGAGGGCCUGAAUUUCAAC-3′ (SEQ ID NO: 1016) 3′-CUUUAGGACCUCCCGGACUUAAAGUUG-5′ (SEQ ID NO: 224) AAT-582 Target: 5′-GAAATCCTGGAGGGCCTGAATTTCAAC-3′ (SEQ ID NO: 422) 5′-AAAUCCUGGAGGGCCUGAAUUUCAACC-3′ (SEQ ID NO: 1017) 3′-UUUAGGACCUCCCGGACUUAAAGUUGG-5′ (SEQ ID NO: 225) AAT-583 Target: 5′-AAATCCTGGAGGGCCTGAATTTCAACC-3′ (SEQ ID NO: 423) 5′-AUCCUGGAGGGCCUGAAUUUCAACCUC-3′ (SEQ ID NO: 1018) 3′-UAGGACCUCCCGGACUUAAAGUUGGAG-5′ (SEQ ID NO: 226) AAT-585 Target: 5′-ATCCTGGAGGGCCTGAATTTCAACCTC-3′ (SEQ ID NO: 424) 5′-UCCUGGAGGGCCUGAAUUUCAACCUCA-3′ (SEQ ID NO: 1019) 3′-AGGACCUCCCGGACUUAAAGUUGGAGU-5′ (SEQ ID NO: 227) AAT-586 Target: 5′-TCCTGGAGGGCCTGAATTTCAACCTCA-3′ (SEQ ID NO: 425) 5′-CCUGGAGGGCCUGAAUUUCAACCUCAC-3′ (SEQ ID NO: 1020) 3′-GGACCUCCCGGACUUAAAGUUGGAGUG-5′ (SEQ ID NO: 228) AAT-587 Target: 5′-CCTGGAGGGCCTGAATTTCAACCTCAC-3′ (SEQ ID NO: 426) 5′-UCCAUGAAGGCUUCCAGGAACUCCUCC-3′ (SEQ ID NO: 1021) 3′-AGGUACUUCCGAAGGUCCUUGAGGAGG-5′ (SEQ ID NO: 229) AAT-634 Target: 5′-TCCATGAAGGCTTCCAGGAACTCCTCC-3′ (SEQ ID NO: 427) 5′-AUGAAGGCUUCCAGGAACUCCUCCGUA-3′ (SEQ ID NO: 1022) 3′-UACUUCCGAAGGUCCUUGAGGAGGCAU-5′ (SEQ ID NO: 230) AAT-637 Target: 5′-ATGAAGGCTTCCAGGAACTCCTCCGTA-3′ (SEQ ID NO: 428) 5′-UGAAGGCUUCCAGGAACUCCUCCGUAC-3′ (SEQ ID NO: 1023) 3′-ACUUCCGAAGGUCCUUGAGGAGGCAUG-5′ (SEQ ID NO: 231) AAT-638 Target: 5′-TGAAGGCTTCCAGGAACTCCTCCGTAC-3′ (SEQ ID NO: 429) 5′-CCAGCCAGACAGCCAGCUCCAGCUGAC-3′ (SEQ ID NO: 1024) 3′-GGUCGGUCUGUCGGUCGAGGUCGACUG-5′ (SEQ ID NO: 232) AAT-671 Target: 5′-CCAGCCAGACAGCCAGCTCCAGCTGAC-3′ (SEQ ID NO: 430) 5′-AGCCAGACAGCCAGCUCCAGCUGACCA-3′ (SEQ ID NO: 1025) 3′-UCGGUCUGUCGGUCGAGGUCGACUGGU-5′ (SEQ ID NO: 233) AAT-673 Target: 5′-AGCCAGACAGCCAGCTCCAGCTGACCA-3′ (SEQ ID NO: 431) 5′-CCAGACAGCCAGCUCCAGCUGACCACC-3′ (SEQ ID NO: 1026) 3′-GGUCUGUCGGUCGAGGUCGACUGGUGG-5′ (SEQ ID NO: 234) AAT-675 Target: 5′-CCAGACAGCCAGCTCCAGCTGACCACC-3′ (SEQ ID NO: 432) 5′-CAGACAGCCAGCUCCAGCUGACCACCG-3′ (SEQ ID NO: 1027) 3′-GUCUGUCGGUCGAGGUCGACUGGUGGC-5′ (SEQ ID NO: 235) AAT-676 Target: 5′-CAGACAGCCAGCTCCAGCTGACCACCG-3′ (SEQ ID NO: 433) 5′-GCUAGUGGAUAAGUUUUUGGAGGAUGU-3′ (SEQ ID NO: 1028) 3′-CGAUCACCUAUUCAAAAACCUCCUACA-5′ (SEQ ID NO: 236) AAT-734 Target: 5′-GCTAGTGGATAAGTTTTTGGAGGATGT-3′ (SEQ ID NO: 434) 5′-CUAGUGGAUAAGUUUUUGGAGGAUGUU-3′ (SEQ ID NO: 1029) 3′-GAUCACCUAUUCAAAAACCUCCUACAA-5′ (SEQ ID NO: 237) AAT-735 Target: 5′-CTAGTGGATAAGTTTTTGGAGGATGTT-3′ (SEQ ID NO: 435) 5′-UAGUGGAUAAGUUUUUGGAGGAUGUUA-3′ (SEQ ID NO: 1030) 3′-AUCACCUAUUCAAAAACCUCCUACAAU-5′ (SEQ ID NO: 238) AAT-736 Target: 5′-TAGTGGATAAGTTTTTGGAGGATGTTA-3′ (SEQ ID NO: 436) 5′-AGUGGAUAAGUUUUUGGAGGAUGUUAA-3′ (SEQ ID NO: 1031) 3′-UCACCUAUUCAAAAACCUCCUACAAUU-5′ (SEQ ID NO: 239) AAT-737 Target: 5′-AGTGGATAAGTTTTTGGAGGATGTTAA-3′ (SEQ ID NO: 437) 5′-GUGGAUAAGUUUUUGGAGGAUGUUAAA-3′ (SEQ ID NO: 1032) 3′-CACCUAUUCAAAAACCUCCUACAAUUU-5′ (SEQ ID NO: 240) AAT-738 Target: 5′-GTGGATAAGTTTTTGGAGGATGTTAAA-3′ (SEQ ID NO: 438) 5′-UGGAUAAGUUUUUGGAGGAUGUUAAAA-3′ (SEQ ID NO: 1033) 3′-ACCUAUUCAAAAACCUCCUACAAUUUU-5′ (SEQ ID NO: 241) AAT-739 Target: 5′-TGGATAAGTTTTTGGAGGATGTTAAAA-3′ (SEQ ID NO: 439) 5′-GGAUAAGUUUUUGGAGGAUGUUAAAAA-3′ (SEQ ID NO: 1034) 3′-CCUAUUCAAAAACCUCCUACAAUUUUU-5′ (SEQ ID NO: 242) AAT-740 Target: 5′-GGATAAGTTTTTGGAGGATGTTAAAAA-3′ (SEQ ID NO: 440) 5′-GUUGUACCACUCAGAAGCCUUCACUGU-3′ (SEQ ID NO: 1035) 3′-CAACAUGGUGAGUCUUCGGAAGUGACA-5′ (SEQ ID NO: 243) AAT-767 Target: 5′-GTTGTACCACTCAGAAGCCTTCACTGT-3′ (SEQ ID NO: 441) 5′-UUGUACCACUCAGAAGCCUUCACUGUC-3′ (SEQ ID NO: 1036) 3′-AACAUGGUGAGUCUUCGGAAGUGACAG-5′ (SEQ ID NO: 244) AAT-768 Target: 5′-TTGTACCACTCAGAAGCCTTCACTGTC-3′ (SEQ ID NO: 442) 5′-GUACUCAAGGGAAAAUUGUGGAUUUGG-3′ (SEQ ID NO: 1037) 3′-CAUGAGUUCCCUUUUAACACCUAAACC-5′ (SEQ ID NO: 245) AAT-850 Target: 5′-GTACTCAAGGGAAAATTGTGGATTTGG-3′ (SEQ ID NO: 443) 5′-UACUCAAGGGAAAAUUGUGGAUUUGGU-3′ (SEQ ID NO: 1038) 3′-AUGAGUUCCCUUUUAACACCUAAACCA-5′ (SEQ ID NO: 246) AAT-851 Target: 5′-TACTCAAGGGAAAATTGTGGATTTGGT-3′ (SEQ ID NO: 444) 5′-ACUCAAGGGAAAAUUGUGGAUUUGGUC-3′ (SEQ ID NO: 1039) 3′-UGAGUUCCCUUUUAACACCUAAACCAG-5′ (SEQ ID NO: 247) AAT-852 Target: 5′-ACTCAAGGGAAAATTGTGGATTTGGTC-3′ (SEQ ID NO: 445) 5′-CUCAAGGGAAAAUUGUGGAUUUGGUCA-3′ (SEQ ID NO: 1040) 3′-GAGUUCCCUUUUAACACCUAAACCAGU-5′ (SEQ ID NO: 248) AAT-853 Target: 5′-CTCAAGGGAAAATTGTGGATTTGGTCA-3′ (SEQ ID NO: 446) 5′-UCAAGGGAAAAUUGUGGAUUUGGUCAA-3′ (SEQ ID NO: 1041) 3′-AGUUCCCUUUUAACACCUAAACCAGUU-5′ (SEQ ID NO: 249) AAT-854 Target: 5′-TCAAGGGAAAATTGTGGATTTGGTCAA-3′ (SEQ ID NO: 447) 5′-CAAGGGAAAAUUGUGGAUUUGGUCAAG-3′ (SEQ ID NO: 1042) 3′-GUUCCCUUUUAACACCUAAACCAGUUC-5′ (SEQ ID NO: 250) AAT-855 Target: 5′-CAAGGGAAAATTGTGGATTTGGTCAAG-3′ (SEQ ID NO: 448) 5′-AAGGGAAAAUUGUGGAUUUGGUCAAGG-3′ (SEQ ID NO: 1043) 3′-UUCCCUUUUAACACCUAAACCAGUUCC-5′ (SEQ ID NO: 251) AAT-856 Target: 5′-AAGGGAAAATTGTGGATTTGGTCAAGG-3′ (SEQ ID NO: 449) 5′-AGGGAAAAUUGUGGAUUUGGUCAAGGA-3′ (SEQ ID NO: 1044) 3′-UCCCUUUUAACACCUAAACCAGUUCCU-5′ (SEQ ID NO: 252) AAT-857 Target: 5′-AGGGAAAATTGTGGATTTGGTCAAGGA-3′ (SEQ ID NO: 450) 5′-GGGAAAAUUGUGGAUUUGGUCAAGGAG-3′ (SEQ ID NO: 1045) 3′-CCCUUUUAACACCUAAACCAGUUCCUC-5′ (SEQ ID NO: 253) AAT-858 Target: 5′-GGGAAAATTGTGGATTTGGTCAAGGAG-3′ (SEQ ID NO: 451) 5′-GGAAAAUUGUGGAUUUGGUCAAGGAGC-3′ (SEQ ID NO: 1046) 3′-CCUUUUAACACCUAAACCAGUUCCUCG-5′ (SEQ ID NO: 254) AAT-859 Target: 5′-GGAAAATTGTGGATTTGGTCAAGGAGC-3′ (SEQ ID NO: 452) 5′-GAAAAUUGUGGAUUUGGUCAAGGAGCU-3′ (SEQ ID NO: 1047) 3′-CUUUUAACACCUAAACCAGUUCCUCGA-5′ (SEQ ID NO: 255) AAT-860 Target: 5′-GAAAATTGTGGATTTGGTCAAGGAGCT-3′ (SEQ ID NO: 453) 5′-AAAAUUGUGGAUUUGGUCAAGGAGCUU-3′ (SEQ ID NO: 1048) 3′-UUUUAACACCUAAACCAGUUCCUCGAA-5′ (SEQ ID NO: 256) AAT-861 Target: 5′-AAAATTGTGGATTTGGTCAAGGAGCTT-3′ (SEQ ID NO: 454) 5′-AAAUUGUGGAUUUGGUCAAGGAGCUUG-3′ (SEQ ID NO: 1049) 3′-UUUAACACCUAAACCAGUUCCUCGAAC-5′ (SEQ ID NO: 257) AAT-862 Target: 5′-AAATTGTGGATTTGGTCAAGGAGCTTG-3′ (SEQ ID NO: 455) 5′-AAUUGUGGAUUUGGUCAAGGAGCUUGA-3′ (SEQ ID NO: 1050) 3′-UUAACACCUAAACCAGUUCCUCGAACU-5′ (SEQ ID NO: 258) AAT-863 Target: 5′-AATTGTGGATTTGGTCAAGGAGCTTGA-3′ (SEQ ID NO: 456) 5′-AUUGUGGAUUUGGUCAAGGAGCUUGAC-3′ (SEQ ID NO: 1051) 3′-UAACACCUAAACCAGUUCCUCGAACUG-5′ (SEQ ID NO: 259) AAT-864 Target: 5′-ATTGTGGATTTGGTCAAGGAGCTTGAC-3′ (SEQ ID NO: 457) 5′-UUGUGGAUUUGGUCAAGGAGCUUGACA-3′ (SEQ ID NO: 1052) 3′-AACACCUAAACCAGUUCCUCGAACUGU-5′ (SEQ ID NO: 260) AAT-865 Target: 5′-TTGTGGATTTGGTCAAGGAGCTTGACA-3′ (SEQ ID NO: 458) 5′-UGUGGAUUUGGUCAAGGAGCUUGACAG-3′ (SEQ ID NO: 1053) 3′-ACACCUAAACCAGUUCCUCGAACUGUC-5′ (SEQ ID NO: 261) AAT-866 Target: 5′-TGTGGATTTGGTCAAGGAGCTTGACAG-3′ (SEQ ID NO: 459) 5′-GUGGAUUUGGUCAAGGAGCUUGACAGA-3′ (SEQ ID NO: 1054) 3′-CACCUAAACCAGUUCCUCGAACUGUCU-5′ (SEQ ID NO: 262) AAT-867 Target: 5′-GTGGATTTGGTCAAGGAGCTTGACAGA-3′ (SEQ ID NO: 460) 5′-UGGAUUUGGUCAAGGAGCUUGACAGAG-3′ (SEQ ID NO: 1055) 3′-ACCUAAACCAGUUCCUCGAACUGUCUC-5′ (SEQ ID NO: 263) AAT-868 Target: 5′-TGGATTTGGTCAAGGAGCTTGACAGAG-3′ (SEQ ID NO: 461) 5′-GGAUUUGGUCAAGGAGCUUGACAGAGA-3′ (SEQ ID NO: 1056) 3′-CCUAAACCAGUUCCUCGAACUGUCUCU-5′ (SEQ ID NO: 264) AAT-869 Target: 5′-GGATTTGGTCAAGGAGCTTGACAGAGA-3′ (SEQ ID NO: 462) 5′-GAUUUGGUCAAGGAGCUUGACAGAGAC-3′ (SEQ ID NO: 1057) 3′-CUAAACCAGUUCCUCGAACUGUCUCUG-5′ (SEQ ID NO: 265) AAT-870 Target: 5′-GATTTGGTCAAGGAGCTTGACAGAGAC-3′ (SEQ ID NO: 463) 5′-AUUUGGUCAAGGAGCUUGACAGAGACA-3′ (SEQ ID NO: 1058) 3′-UAAACCAGUUCCUCGAACUGUCUCUGU-5′ (SEQ ID NO: 266) AAT-871 Target: 5′-ATTTGGTCAAGGAGCTTGACAGAGACA-3′ (SEQ ID NO: 464) 5′-UUUGGUCAAGGAGCUUGACAGAGACAC-3′ (SEQ ID NO: 1059) 3′-AAACCAGUUCCUCGAACUGUCUCUGUG-5′ (SEQ ID NO: 267) AAT-872 Target: 5′-TTTGGTCAAGGAGCTTGACAGAGACAC-3′ (SEQ ID NO: 465) 5′-CACAGUUUUUGCUCUGGUGAAUUACAU-3′ (SEQ ID NO: 1060) 3′-GUGUCAAAAACGAGACCACUUAAUGUA-5′ (SEQ ID NO: 268) AAT-896 Target: 5′-CACAGTTTTTGCTCTGGTGAATTACAT-3′ (SEQ ID NO: 466) 5′-ACAGUUUUUGCUCUGGUGAAUUACAUC-3′ (SEQ ID NO: 1061) 3′-UGUCAAAAACGAGACCACUUAAUGUAG-5′ (SEQ ID NO: 269) AAT-897 Target: 5′-ACAGTTTTTGCTCTGGTGAATTACATC-3′ (SEQ ID NO: 467) 5′-CAGUUUUUGCUCUGGUGAAUUACAUCU-3′ (SEQ ID NO: 1062) 3′-GUCAAAAACGAGACCACUUAAUGUAGA-5′ (SEQ ID NO: 270) AAT-898 Target: 5′-CAGTTTTTGCTCTGGTGAATTACATCT-3′ (SEQ ID NO: 468) 5′-AGUUUUUGCUCUGGUGAAUUACAUCUU-3′ (SEQ ID NO: 1063) 3′-UCAAAAACGAGACCACUUAAUGUAGAA-5′ (SEQ ID NO: 271) AAT-899 Target: 5′-AGTTTTTGCTCTGGTGAATTACATCTT-3′ (SEQ ID NO: 469) 5′-UUAAAGGCAAAUGGGAGAGACCCUUUG-3′ (SEQ ID NO: 1064) 3′-AAUUUCCGUUUACCCUCUCUGGGAAAC-5′ (SEQ ID NO: 272) AAT-928 Target: 5′-TTAAAGGCAAATGGGAGAGACCCTTTG-3′ (SEQ ID NO: 470) 5′-UAAAGGCAAAUGGGAGAGACCCUUUGA-3′ (SEQ ID NO: 1065) 3′-AUUUCCGUUUACCCUCUCUGGGAAACU-5′ (SEQ ID NO: 273) AAT-929 Target: 5′-TAAAGGCAAATGGGAGAGACCCTTTGA-3′ (SEQ ID NO: 471) 5′-AAAGGCAAAUGGGAGAGACCCUUUGAA-3′ (SEQ ID NO: 1066) 3′-UUUCCGUUUACCCUCUCUGGGAAACUU-5′ (SEQ ID NO: 274) AAT-930 Target: 5′-AAAGGCAAATGGGAGAGACCCTTTGAA-3′ (SEQ ID NO: 472) 5′-AAGGCAAAUGGGAGAGACCCUUUGAAG-3′ (SEQ ID NO: 1067) 3′-UUCCGUUUACCCUCUCUGGGAAACUUC-5′ (SEQ ID NO: 275) AAT-931 Target: 5′-AAGGCAAATGGGAGAGACCCTTTGAAG-3′ (SEQ ID NO: 473) 5′-CGAGGAAGAGGACUUCCACGUGGACCA-3′ (SEQ ID NO: 1068) 3′-GCUCCUUCUCCUGAAGGUGCACCUGGU-5′ (SEQ ID NO: 276) AAT-968 Target: 5′-CGAGGAAGAGGACTTCCACGTGGACCA-3′ (SEQ ID NO: 474) 5′-GAGGAAGAGGACUUCCACGUGGACCAG-3′ (SEQ ID NO: 1069) 3′-CUCCUUCUCCUGAAGGUGCACCUGGUC-5′ (SEQ ID NO: 277) AAT-969 Target: 5′-GAGGAAGAGGACTTCCACGTGGACCAG-3′ (SEQ ID NO: 475) 5′-AGGAAGAGGACUUCCACGUGGACCAGG-3′ (SEQ ID NO: 1070) 3′-UCCUUCUCCUGAAGGUGCACCUGGUCC-5′ (SEQ ID NO: 278) AAT-970 Target: 5′-AGGAAGAGGACTTCCACGTGGACCAGG-3′ (SEQ ID NO: 476) 5′-GGAAGAGGACUUCCACGUGGACCAGGU-3′ (SEQ ID NO: 1071) 3′-CCUUCUCCUGAAGGUGCACCUGGUCCA-5′ (SEQ ID NO: 279) AAT-971 Target: 5′-GGAAGAGGACTTCCACGTGGACCAGGT-3′ (SEQ ID NO: 477) 5′-AAGAGGACUUCCACGUGGACCAGGUGA-3′ (SEQ ID NO: 1072) 3′-UUCUCCUGAAGGUGCACCUGGUCCACU-5′ (SEQ ID NO: 280) AAT-973 Target: 5′-AAGAGGACTTCCACGTGGACCAGGTGA-3′ (SEQ ID NO: 478) 5′-AGAGGACUUCCACGUGGACCAGGUGAC-3′ (SEQ ID NO: 1073) 3′-UCUCCUGAAGGUGCACCUGGUCCACUG-5′ (SEQ ID NO: 281) AAT-974 Target: 5′-AGAGGACTTCCACGTGGACCAGGTGAC-3′ (SEQ ID NO: 479) 5′-AGGACUUCCACGUGGACCAGGUGACCA-3′ (SEQ ID NO: 1074) 3′-UCCUGAAGGUGCACCUGGUCCACUGGU-5′ (SEQ ID NO: 282) AAT-976 Target: 5′-AGGACTTCCACGTGGACCAGGTGACCA-3′ (SEQ ID NO: 480) 5′-GCGUUUAGGCAUGUUUAACAUCCAGCA-3′ (SEQ ID NO: 1075) 3′-CGCAAAUCCGUACAAAUUGUAGGUCGU-5′ (SEQ ID NO: 283) AAT-1025 Target: 5′-GCGTTTAGGCATGTTTAACATCCAGCA-3′ (SEQ ID NO: 481) 5′-CGUUUAGGCAUGUUUAACAUCCAGCAC-3′ (SEQ ID NO: 1076) 3′-GCAAAUCCGUACAAAUUGUAGGUCGUG-5′ (SEQ ID NO: 284) AAT-1026 Target: 5′-CGTTTAGGCATGTTTAACATCCAGCAC-3′ (SEQ ID NO: 482) 5′-AAGCUGUCCAGCUGGGUGCUGCUGAUG-3′ (SEQ ID NO: 1077) 3′-UUCGACAGGUCGACCCACGACGACUAC-5′ (SEQ ID NO: 285) AAT-1059 Target: 5′-AAGCTGTCCAGCTGGGTGCTGCTGATG-3′ (SEQ ID NO: 483) 5′-AGCUGUCCAGCUGGGUGCUGCUGAUGA-3′ (SEQ ID NO: 1078) 3′-UCGACAGGUCGACCCACGACGACUACU-5′ (SEQ ID NO: 286) AAT-1060 Target: 5′-AGCTGTCCAGCTGGGTGCTGCTGATGA-3′ (SEQ ID NO: 484) 5′-GGCAAUGCCACCGCCAUCUUCUUCCUG-3′ (SEQ ID NO: 1079) 3′-CCGUUACGGUGGCGGUAGAAGAAGGAC-5′ (SEQ ID NO: 287) AAT-1095 Target: 5′-GGCAATGCCACCGCCATCTTCTTCCTG-3′ (SEQ ID NO: 485) 5′-GCAAUGCCACCGCCAUCUUCUUCCUGC-3′ (SEQ ID NO: 1080) 3′-CGUUACGGUGGCGGUAGAAGAAGGACG-5′ (SEQ ID NO: 288) AAT-1096 Target: 5′-GCAATGCCACCGCCATCTTCTTCCTGC-3′ (SEQ ID NO: 486) 5′-UGCCACCGCCAUCUUCUUCCUGCCUGA-3′ (SEQ ID NO: 1081) 3′-ACGGUGGCGGUAGAAGAAGGACGGACU-5′ (SEQ ID NO: 289) AAT-1100 Target: 5′-TGCCACCGCCATCTTCTTCCTGCCTGA-3′ (SEQ ID NO: 487) 5′-GCCACCGCCAUCUUCUUCCUGCCUGAU-3′ (SEQ ID NO: 1082) 3′-CGGUGGCGGUAGAAGAAGGACGGACUA-5′ (SEQ ID NO: 290) AAT-1101 Target: 5′-GCCACCGCCATCTTCTTCCTGCCTGAT-3′ (SEQ ID NO: 488) 5′-CCACCGCCAUCUUCUUCCUGCCUGAUG-3′ (SEQ ID NO: 1083) 3′-GGUGGCGGUAGAAGAAGGACGGACUAC-5′ (SEQ ID NO: 291) AAT-1102 Target: 5′-CCACCGCCATCTTCTTCCTGCCTGATG-3′ (SEQ ID NO: 489) 5′-CACCGCCAUCUUCUUCCUGCCUGAUGA-3′ (SEQ ID NO: 1084) 3′-GUGGCGGUAGAAGAAGGACGGACUACU-5′ (SEQ ID NO: 292) AAT-1103 Target: 5′-CACCGCCATCTTCTTCCTGCCTGATGA-3′ (SEQ ID NO: 490) 5′-ACCGCCAUCUUCUUCCUGCCUGAUGAG-3′ (SEQ ID NO: 1085) 3′-UGGCGGUAGAAGAAGGACGGACUACUC-5′ (SEQ ID NO: 293) AAT-1104 Target: 5′-ACCGCCATCTTCTTCCTGCCTGATGAG-3′ (SEQ ID NO: 491) 5′-CCGCCAUCUUCUUCCUGCCUGAUGAGG-3′ (SEQ ID NO: 1086) 3′-GGCGGUAGAAGAAGGACGGACUACUCC-5′ (SEQ ID NO: 294) AAT-1105 Target: 5′-CCGCCATCTTCTTCCTGCCTGATGAGG-3′ (SEQ ID NO: 492) 5′-CCAUCUUCUUCCUGCCUGAUGAGGGGA-3′ (SEQ ID NO: 1087) 3′-GGUAGAAGAAGGACGGACUACUCCCCU-5′ (SEQ ID NO: 295) AAT-1108 Target: 5′-CCATCTTCTTCCTGCCTGATGAGGGGA-3′ (SEQ ID NO: 493) 5′-UUCUUCCUGCCUGAUGAGGGGAAACUA-3′ (SEQ ID NO: 1088) 3′-AAGAAGGACGGACUACUCCCCUUUGAU-5′ (SEQ ID NO: 296) AAT-1113 Target: 5′-TTCTTCCTGCCTGATGAGGGGAAACTA-3′ (SEQ ID NO: 494) 5′-UCUUCCUGCCUGAUGAGGGGAAACUAC-3′ (SEQ ID NO: 1089) 3′-AGAAGGACGGACUACUCCCCUUUGAUG-5′ (SEQ ID NO: 297) AAT-1114 Target: 5′-TCTTCCTGCCTGATGAGGGGAAACTAC-3′ (SEQ ID NO: 495) 5′-CUUCCUGCCUGAUGAGGGGAAACUACA-3′ (SEQ ID NO: 1090) 3′-GAAGGACGGACUACUCCCCUUUGAUGU-5′ (SEQ ID NO: 298) AAT-1115 Target: 5′-CTTCCTGCCTGATGAGGGGAAACTACA-3′ (SEQ ID NO: 496) 5′-UUCCUGCCUGAUGAGGGGAAACUACAG-3′ (SEQ ID NO: 1091) 3′-AAGGACGGACUACUCCCCUUUGAUGUC-5′ (SEQ ID NO: 299) AAT-1116 Target: 5′-TTCCTGCCTGATGAGGGGAAACTACAG-3′ (SEQ ID NO: 497) 5′-UCCUGCCUGAUGAGGGGAAACUACAGC-3′ (SEQ ID NO: 1092) 3′-AGGACGGACUACUCCCCUUUGAUGUCG-5′ (SEQ ID NO: 300) AAT-1117 Target: 5′-TCCTGCCTGATGAGGGGAAACTACAGC-3′ (SEQ ID NO: 498) 5′-CCUGCCUGAUGAGGGGAAACUACAGCA-3′ (SEQ ID NO: 1093) 3′-GGACGGACUACUCCCCUUUGAUGUCGU-5′ (SEQ ID NO: 301) AAT-1118 Target: 5′-CCTGCCTGATGAGGGGAAACTACAGCA-3′ (SEQ ID NO: 499) 5′-ACAGCACCUGGAAAAUGAACUCACCCA-3′ (SEQ ID NO: 1094) 3′-UGUCGUGGACCUUUUACUUGAGUGGGU-5′ (SEQ ID NO: 302) AAT-1139 Target: 5′-ACAGCACCTGGAAAATGAACTCACCCA-3′ (SEQ ID NO: 500) 5′-CAGCACCUGGAAAAUGAACUCACCCAC-3′ (SEQ ID NO: 1095) 3′-GUCGUGGACCUUUUACUUGAGUGGGUG-5′ (SEQ ID NO: 303) AAT-1140 Target: 5′-CAGCACCTGGAAAATGAACTCACCCAC-3′ (SEQ ID NO: 501) 5′-AGCACCUGGAAAAUGAACUCACCCACG-3′ (SEQ ID NO: 1096) 3′-UCGUGGACCUUUUACUUGAGUGGGUGC-5′ (SEQ ID NO: 304) AAT-1141 Target: 5′-AGCACCTGGAAAATGAACTCACCCACG-3′ (SEQ ID NO: 502) 5′-GCACCUGGAAAAUGAACUCACCCACGA-3′ (SEQ ID NO: 1097) 3′-CGUGGACCUUUUACUUGAGUGGGUGCU-5′ (SEQ ID NO: 305) AAT-1142 Target: 5′-GCACCTGGAAAATGAACTCACCCACGA-3′ (SEQ ID NO: 503) 5′-CACCUGGAAAAUGAACUCACCCACGAU-3′ (SEQ ID NO: 1098) 3′-GUGGACCUUUUACUUGAGUGGGUGCUA-5′ (SEQ ID NO: 306) AAT-1143 Target: 5′-CACCTGGAAAATGAACTCACCCACGAT-3′ (SEQ ID NO: 504) 5′-CGAUAUCAUCACCAAGUUCCUGGAAAA-3′ (SEQ ID NO: 1099) 3′-GCUAUAGUAGUGGUUCAAGGACCUUUU-5′ (SEQ ID NO: 307) AAT-1166 Target: 5′-CGATATCATCACCAAGTTCCTGGAAAA-3′ (SEQ ID NO: 505) 5′-GAUAUCAUCACCAAGUUCCUGGAAAAU-3′ (SEQ ID NO: 1100) 3′-CUAUAGUAGUGGUUCAAGGACCUUUUA-5′ (SEQ ID NO: 308) AAT-1167 Target: 5′-GATATCATCACCAAGTTCCTGGAAAAT-3′ (SEQ ID NO: 506) 5′-AUAUCAUCACCAAGUUCCUGGAAAAUG-3′ (SEQ ID NO: 1101) 3′-UAUAGUAGUGGUUCAAGGACCUUUUAC-5′ (SEQ ID NO: 309) AAT-1168 Target: 5′-ATATCATCACCAAGTTCCTGGAAAATG-3′ (SEQ ID NO: 507) 5′-UAUCAUCACCAAGUUCCUGGAAAAUGA-3′ (SEQ ID NO: 1102) 3′-AUAGUAGUGGUUCAAGGACCUUUUACU-5′ (SEQ ID NO: 310) AAT-1169 Target: 5′-TATCATCACCAAGTTCCTGGAAAATGA-3′ (SEQ ID NO: 508) 5′-AUCAUCACCAAGUUCCUGGAAAAUGAA-3′ (SEQ ID NO: 1103) 3′-UAGUAGUGGUUCAAGGACCUUUUACUU-5′ (SEQ ID NO: 311) AAT-1170 Target: 5′-ATCATCACCAAGTTCCTGGAAAATGAA-3′ (SEQ ID NO: 509) 5′-UCAUCACCAAGUUCCUGGAAAAUGAAG-3′ (SEQ ID NO: 1104) 3′-AGUAGUGGUUCAAGGACCUUUUACUUC-5′ (SEQ ID NO: 312) AAT-1171 Target: 5′-TCATCACCAAGTTCCTGGAAAATGAAG-3′ (SEQ ID NO: 510) 5′-CAUCACCAAGUUCCUGGAAAAUGAAGA-3′ (SEQ ID NO: 1105) 3′-GUAGUGGUUCAAGGACCUUUUACUUCU-5′ (SEQ ID NO: 313) AAT-1172 Target: 5′-CATCACCAAGTTCCTGGAAAATGAAGA-3′ (SEQ ID NO: 511) 5′-AUCACCAAGUUCCUGGAAAAUGAAGAC-3′ (SEQ ID NO: 1106) 3′-UAGUGGUUCAAGGACCUUUUACUUCUG-5′ (SEQ ID NO: 314) AAT-1173 Target: 5′-ATCACCAAGTTCCTGGAAAATGAAGAC-3′ (SEQ ID NO: 512) 5′-UCACCAAGUUCCUGGAAAAUGAAGACA-3′ (SEQ ID NO: 1107) 3′-AGUGGUUCAAGGACCUUUUACUUCUGU-5′ (SEQ ID NO: 315) AAT-1174 Target: 5′-TCACCAAGTTCCTGGAAAATGAAGACA-3′ (SEQ ID NO: 513) 5′-CACCAAGUUCCUGGAAAAUGAAGACAG-3′ (SEQ ID NO: 1108) 3′-GUGGUUCAAGGACCUUUUACUUCUGUC-5′ (SEQ ID NO: 316) AAT-1175 Target: 5′-CACCAAGTTCCTGGAAAATGAAGACAG-3′ (SEQ ID NO: 514) 5′-UAAGGUCUUCAGCAAUGGGGCUGACCU-3′ (SEQ ID NO: 1109) 3′-AUUCCAGAAGUCGUUACCCCGACUGGA-5′ (SEQ ID NO: 317) AAT-1286 Target: 5′-TAAGGTCTTCAGCAATGGGGCTGACCT-3′ (SEQ ID NO: 515) 5′-AGCAAUGGGGCUGACCUCUCCGGGGUC-3′ (SEQ ID NO: 1110) 3′-UCGUUACCCCGACUGGAGAGGCCCCAG-5′ (SEQ ID NO: 318) AAT-1296 Target: 5′-AGCAATGGGGCTGACCTCTCCGGGGTC-3′ (SEQ ID NO: 516) 5′-GCAAUGGGGCUGACCUCUCCGGGGUCA-3′ (SEQ ID NO: 1111) 3′-CGUUACCCCGACUGGAGAGGCCCCAGU-5′ (SEQ ID NO: 319) AAT-1297 Target: 5′-GCAATGGGGCTGACCTCTCCGGGGTCA-3′ (SEQ ID NO: 517) 5′-CAAUGGGGCUGACCUCUCCGGGGUCAC-3′ (SEQ ID NO: 1112) 3′-GUUACCCCGACUGGAGAGGCCCCAGUG-5′ (SEQ ID NO: 320) AAT-1298 Target: 5′-CAATGGGGCTGACCTCTCCGGGGTCAC-3′ (SEQ ID NO: 518) 5′-CAGAGGAGGCACCCCUGAAGCUCUCCA-3′ (SEQ ID NO: 1113) 3′-GUCUCCUCCGUGGGGACUUCGAGAGGU-5′ (SEQ ID NO: 321) AAT-1324 Target: 5′-CAGAGGAGGCACCCCTGAAGCTCTCCA-3′ (SEQ ID NO: 519) 5′-GAGGAGGCACCCCUGAAGCUCUCCAAG-3′ (SEQ ID NO: 1114) 3′-CUCCUCCGUGGGGACUUCGAGAGGUUC-5′ (SEQ ID NO: 322) AAT-1326 Target: 5′-GAGGAGGCACCCCTGAAGCTCTCCAAG-3′ (SEQ ID NO: 520) 5′-CCCUGAAGCUCUCCAAGGCCGUGCAUA-3′ (SEQ ID NO: 1115) 3′-GGGACUUCGAGAGGUUCCGGCACGUAU-5′ (SEQ ID NO: 323) AAT-1336 Target: 5′-CCCTGAAGCTCTCCAAGGCCGTGCATA-3′ (SEQ ID NO: 521) 5′-GCCGUGCAUAAGGCUGUGCUGACCAUC-3′ (SEQ ID NO: 1116) 3′-CGGCACGUAUUCCGACACGACUGGUAG-5′ (SEQ ID NO: 324) AAT-1353 Target: 5′-GCCGTGCATAAGGCTGTGCTGACCATC-3′ (SEQ ID NO: 522) 5′-CCGUGCAUAAGGCUGUGCUGACCAUCG-3′ (SEQ ID NO: 1117) 3′-GGCACGUAUUCCGACACGACUGGUAGC-5′ (SEQ ID NO: 325) AAT-1354 Target: 5′-CCGTGCATAAGGCTGTGCTGACCATCG-3′ (SEQ ID NO: 523) 5′-CGUGCAUAAGGCUGUGCUGACCAUCGA-3′ (SEQ ID NO: 1118) 3′-GCACGUAUUCCGACACGACUGGUAGCU-5′ (SEQ ID NO: 326) AAT-1355 Target: 5′-CGTGCATAAGGCTGTGCTGACCATCGA-3′ (SEQ ID NO: 524) 5′-GUGCAUAAGGCUGUGCUGACCAUCGAC-3′ (SEQ ID NO: 1119) 3′-CACGUAUUCCGACACGACUGGUAGCUG-5′ (SEQ ID NO: 327) AAT-1356 Target: 5′-GTGCATAAGGCTGTGCTGACCATCGAC-3′ (SEQ ID NO: 525) 5′-UGCAUAAGGCUGUGCUGACCAUCGACG-3′ (SEQ ID NO: 1120) 3′-ACGUAUUCCGACACGACUGGUAGCUGC-5′ (SEQ ID NO: 328) AAT-1357 Target: 5′-TGCATAAGGCTGTGCTGACCATCGACG-3′ (SEQ ID NO: 526) 5′-GCAUAAGGCUGUGCUGACCAUCGACGA-3′ (SEQ ID NO: 1121) 3′-CGUAUUCCGACACGACUGGUAGCUGCU-5′ (SEQ ID NO: 329) AAT-1358 Target: 5′-GCATAAGGCTGTGCTGACCATCGACGA-3′ (SEQ ID NO: 527) 5′-CAUAAGGCUGUGCUGACCAUCGACGAG-3′ (SEQ ID NO: 1122) 3′-GUAUUCCGACACGACUGGUAGCUGCUC-5′ (SEQ ID NO: 330) AAT-1359 Target: 5′-CATAAGGCTGTGCTGACCATCGACGAG-3′ (SEQ ID NO: 528) 5′-AUAAGGCUGUGCUGACCAUCGACGAGA-3′ (SEQ ID NO: 1123) 3′-UAUUCCGACACGACUGGUAGCUGCUCU-5′ (SEQ ID NO: 331) AAT-1360 Target: 5′-ATAAGGCTGTGCTGACCATCGACGAGA-3′ (SEQ ID NO: 529) 5′-UAAGGCUGUGCUGACCAUCGACGAGAA-3′ (SEQ ID NO: 1124) 3′-AUUCCGACACGACUGGUAGCUGCUCUU-5′ (SEQ ID NO: 332) AAT-1361 Target: 5′-TAAGGCTGTGCTGACCATCGACGAGAA-3′ (SEQ ID NO: 530) 5′-GGACUGAAGCUGCUGGGGCCAUGUUUU-3′ (SEQ ID NO: 1125) 3′-CCUGACUUCGACGACCCCGGUACAAAA-5′ (SEQ ID NO: 333) AAT-1390 Target: 5′-GGACTGAAGCTGCTGGGGCCATGTTTT-3′ (SEQ ID NO: 531) 5′-GACUGAAGCUGCUGGGGCCAUGUUUUU-3′ (SEQ ID NO: 1126) 3′-CUGACUUCGACGACCCCGGUACAAAAA-5′ (SEQ ID NO: 334) AAT-1391 Target: 5′-GACTGAAGCTGCTGGGGCCATGTTTTT-3′ (SEQ ID NO: 532) 5′-ACUGAAGCUGCUGGGGCCAUGUUUUUA-3′ (SEQ ID NO: 1127) 3′-UGACUUCGACGACCCCGGUACAAAAAU-5′ (SEQ ID NO: 335) AAT-1392 Target: 5′-ACTGAAGCTGCTGGGGCCATGTTTTTA-3′ (SEQ ID NO: 533) 5′-CUGAAGCUGCUGGGGCCAUGUUUUUAG-3′ (SEQ ID NO: 1128) 3′-GACUUCGACGACCCCGGUACAAAAAUC-5′ (SEQ ID NO: 336) AAT-1393 Target: 5′-CTGAAGCTGCTGGGGCCATGTTTTTAG-3′ (SEQ ID NO: 534) 5′-UGAAGCUGCUGGGGCCAUGUUUUUAGA-3′ (SEQ ID NO: 1129) 3′-ACUUCGACGACCCCGGUACAAAAAUCU-5′ (SEQ ID NO: 337) AAT-1394 Target: 5′-TGAAGCTGCTGGGGCCATGTTTTTAGA-3′ (SEQ ID NO: 535) 5′-GAAGCUGCUGGGGCCAUGUUUUUAGAG-3′ (SEQ ID NO: 1130) 3′-CUUCGACGACCCCGGUACAAAAAUCUC-5′ (SEQ ID NO: 338) AAT-1395 Target: 5′-GAAGCTGCTGGGGCCATGTTTTTAGAG-3′ (SEQ ID NO: 536) 5′-GGGCCAUGUUUUUAGAGGCCAUACCCA-3′ (SEQ ID NO: 1131) 3′-CCCGGUACAAAAAUCUCCGGUAUGGGU-5′ (SEQ ID NO: 339) AAT-1405 Target: 5′-GGGCCATGTTTTTAGAGGCCATACCCA-3′ (SEQ ID NO: 537) 5′-GGCCAUGUUUUUAGAGGCCAUACCCAU-3′ (SEQ ID NO: 1132) 3′-CCGGUACAAAAAUCUCCGGUAUGGGUA-5′ (SEQ ID NO: 340) AAT-1406 Target: 5′-GGCCATGTTTTTAGAGGCCATACCCAT-3′ (SEQ ID NO: 538) 5′-GCCAUGUUUUUAGAGGCCAUACCCAUG-3′ (SEQ ID NO: 1133) 3′-CGGUACAAAAAUCUCCGGUAUGGGUAC-5′ (SEQ ID NO: 341) AAT-1407 Target: 5′-GCCATGTTTTTAGAGGCCATACCCATG-3′ (SEQ ID NO: 539) 5′-CCAUGUUUUUAGAGGCCAUACCCAUGU-3′ (SEQ ID NO: 1134) 3′-GGUACAAAAAUCUCCGGUAUGGGUACA-5′ (SEQ ID NO: 342) AAT-1408 Target: 5′-CCATGTTTTTAGAGGCCATACCCATGT-3′ (SEQ ID NO: 540) 5′-CAUGUUUUUAGAGGCCAUACCCAUGUC-3′ (SEQ ID NO: 1135) 3′-GUACAAAAAUCUCCGGUAUGGGUACAG-5′ (SEQ ID NO: 343) AAT-1409 Target: 5′-CATGTTTTTAGAGGCCATACCCATGTC-3′ (SEQ ID NO: 541) 5′-AUGUUUUUAGAGGCCAUACCCAUGUCU-3′ (SEQ ID NO: 1136) 3′-UACAAAAAUCUCCGGUAUGGGUACAGA-5′ (SEQ ID NO: 344) AAT-1410 Target: 5′-ATGTTTTTAGAGGCCATACCCATGTCT-3′ (SEQ ID NO: 542) 5′-UGUUUUUAGAGGCCAUACCCAUGUCUA-3′ (SEQ ID NO: 1137) 3′-ACAAAAAUCUCCGGUAUGGGUACAGAU-5′ (SEQ ID NO: 345) AAT-1411 Target: 5′-TGTTTTTAGAGGCCATACCCATGTCTA-3′ (SEQ ID NO: 543) 5′-GUUUUUAGAGGCCAUACCCAUGUCUAU-3′ (SEQ ID NO: 1138) 3′-CAAAAAUCUCCGGUAUGGGUACAGAUA-5′ (SEQ ID NO: 346) AAT-1412 Target: 5′-GTTTTTAGAGGCCATACCCATGTCTAT-3′ (SEQ ID NO: 544) 5′-UUUUUAGAGGCCAUACCCAUGUCUAUC-3′ (SEQ ID NO: 1139) 3′-AAAAAUCUCCGGUAUGGGUACAGAUAG-5′ (SEQ ID NO: 347) AAT-1413 Target: 5′-TTTTTAGAGGCCATACCCATGTCTATC-3′ (SEQ ID NO: 545) 5′-UUUUAGAGGCCAUACCCAUGUCUAUCC-3′ (SEQ ID NO: 1140) 3′-AAAAUCUCCGGUAUGGGUACAGAUAGG-5′ (SEQ ID NO: 348) AAT-1414 Target: 5′-TTTTAGAGGCCATACCCATGTCTATCC-3′ (SEQ ID NO: 546) 5′-UUUAGAGGCCAUACCCAUGUCUAUCCC-3′ (SEQ ID NO: 1141) 3′-AAAUCUCCGGUAUGGGUACAGAUAGGG-5′ (SEQ ID NO: 349) AAT-1415 Target: 5′-TTTAGAGGCCATACCCATGTCTATCCC-3′ (SEQ ID NO: 547) 5′-UUAGAGGCCAUACCCAUGUCUAUCCCC-3′ (SEQ ID NO: 1142) 3′-AAUCUCCGGUAUGGGUACAGAUAGGGG-5′ (SEQ ID NO: 350) AAT-1416 Target: 5′-TTAGAGGCCATACCCATGTCTATCCCC-3′ (SEQ ID NO: 548) 5′-AAGUUCAACAAACCCUUUGUCUUCUUA-3′ (SEQ ID NO: 1143) 3′-UUCAAGUUGUUUGGGAAACAGAAGAAU-5′ (SEQ ID NO: 351) AAT-1452 Target: 5′-AAGTTCAACAAACCCTTTGTCTTCTTA-3′ (SEQ ID NO: 549) 5′-AGUUCAACAAACCCUUUGUCUUCUUAA-3′ (SEQ ID NO: 1144) 3′-UCAAGUUGUUUGGGAAACAGAAGAAUU-5′ (SEQ ID NO: 352) AAT-1453 Target: 5′-AGTTCAACAAACCCTTTGTCTTCTTAA-3′ (SEQ ID NO: 550) 5′-GUUCAACAAACCCUUUGUCUUCUUAAU-3′ (SEQ ID NO: 1145) 3′-CAAGUUGUUUGGGAAACAGAAGAAUUA-5′ (SEQ ID NO: 353) AAT-1454 Target: 5′-GTTCAACAAACCCTTTGTCTTCTTAAT-3′ (SEQ ID NO: 551) 5′-UUCAACAAACCCUUUGUCUUCUUAAUG-3′ (SEQ ID NO: 1146) 3′-AAGUUGUUUGGGAAACAGAAGAAUUAC-5′ (SEQ ID NO: 354) AAT-1455 Target: 5′-TTCAACAAACCCTTTGTCTTCTTAATG-3′ (SEQ ID NO: 552) 5′-UCAACAAACCCUUUGUCUUCUUAAUGA-3′ (SEQ ID NO: 1147) 3′-AGUUGUUUGGGAAACAGAAGAAUUACU-5′ (SEQ ID NO: 355) AAT-1456 Target: 5′-TCAACAAACCCTTTGTCTTCTTAATGA-3′ (SEQ ID NO: 553) 5′-CAACAAACCCUUUGUCUUCUUAAUGAU-3′ (SEQ ID NO: 1148) 3′-GUUGUUUGGGAAACAGAAGAAUUACUA-5′ (SEQ ID NO: 356) AAT-1457 Target: 5′-CAACAAACCCTTTGTCTTCTTAATGAT-3′ (SEQ ID NO: 554) 5′-AACAAACCCUUUGUCUUCUUAAUGAUU-3′ (SEQ ID NO: 1149) 3′-UUGUUUGGGAAACAGAAGAAUUACUAA-5′ (SEQ ID NO: 357) AAT-1458 Target: 5′-AACAAACCCTTTGTCTTCTTAATGATT-3′ (SEQ ID NO: 555) 5′-ACAAACCCUUUGUCUUCUUAAUGAUUG-3′ (SEQ ID NO: 1150) 3′-UGUUUGGGAAACAGAAGAAUUACUAAC-5′ (SEQ ID NO: 358) AAT-1459 Target: 5′-ACAAACCCTTTGTCTTCTTAATGATTG-3′ (SEQ ID NO: 556) 5′-CAAACCCUUUGUCUUCUUAAUGAUUGA-3′ (SEQ ID NO: 1151) 3′-GUUUGGGAAACAGAAGAAUUACUAACU-5′ (SEQ ID NO: 359) AAT-1460 Target: 5′-CAAACCCTTTGTCTTCTTAATGATTGA-3′ (SEQ ID NO: 557) 5′-AAAAUACCAAGUCUCCCCUCUUCAUGG-3′ (SEQ ID NO: 1152) 3′-UUUUAUGGUUCAGAGGGGAGAAGUACC-5′ (SEQ ID NO: 360) AAT-1489 Target: 5′-AAAATACCAAGTCTCCCCTCTTCATGG-3′ (SEQ ID NO: 558) 5′-AAAUACCAAGUCUCCCCUCUUCAUGGG-3′ (SEQ ID NO: 1153) 3′-UUUAUGGUUCAGAGGGGAGAAGUACCC-5′ (SEQ ID NO: 361) AAT-1490 Target: 5′-AAATACCAAGTCTCCCCTCTTCATGGG-3′ (SEQ ID NO: 559) 5′-AAUACCAAGUCUCCCCUCUUCAUGGGA-3′ (SEQ ID NO: 1154) 3′-UUAUGGUUCAGAGGGGAGAAGUACCCU-5′ (SEQ ID NO: 362) AAT-1491 Target: 5′-AATACCAAGTCTCCCCTCTTCATGGGA-3′ (SEQ ID NO: 560) 5′-AUACCAAGUCUCCCCUCUUCAUGGGAA-3′ (SEQ ID NO: 1155) 3′-UAUGGUUCAGAGGGGAGAAGUACCCUU-5′ (SEQ ID NO: 363) AAT-1492 Target: 5′-ATACCAAGTCTCCCCTCTTCATGGGAA-3′ (SEQ ID NO: 561) 5′-UACCAAGUCUCCCCUCUUCAUGGGAAA-3′ (SEQ ID NO: 1156) 3′-AUGGUUCAGAGGGGAGAAGUACCCUUU-5′ (SEQ ID NO: 364) AAT-1493 Target: 5′-TACCAAGTCTCCCCTCTTCATGGGAAA-3′ (SEQ ID NO: 562) 5′-ACCAAGUCUCCCCUCUUCAUGGGAAAA-3′ (SEQ ID NO: 1157) 3′-UGGUUCAGAGGGGAGAAGUACCCUUUU-5′ (SEQ ID NO: 365) AAT-1494 Target: 5′-ACCAAGTCTCCCCTCTTCATGGGAAAA-3′ (SEQ ID NO: 563) 5′-CCAAGUCUCCCCUCUUCAUGGGAAAAG-3′ (SEQ ID NO: 1158) 3′-GGUUCAGAGGGGAGAAGUACCCUUUUC-5′ (SEQ ID NO: 366) AAT-1495 Target: 5′-CCAAGTCTCCCCTCTTCATGGGAAAAG-3′ (SEQ ID NO: 564) 5′-CAAGUCUCCCCUCUUCAUGGGAAAAGU-3′ (SEQ ID NO: 1159) 3′-GUUCAGAGGGGAGAAGUACCCUUUUCA-5′ (SEQ ID NO: 367) AAT-1496 Target: 5′-CAAGTCTCCCCTCTTCATGGGAAAAGT-3′ (SEQ ID NO: 565) 5′-AAGUCUCCCCUCUUCAUGGGAAAAGUG-3′ (SEQ ID NO: 1160) 3′-UUCAGAGGGGAGAAGUACCCUUUUCAC-5′ (SEQ ID NO: 368) AAT-1497 Target: 5′-AAGTCTCCCCTCTTCATGGGAAAAGTG-3′ (SEQ ID NO: 566) 5′-GUCUCCCCUCUUCAUGGGAAAAGUGGU-3′ (SEQ ID NO: 1161) 3′-CAGAGGGGAGAAGUACCCUUUUCACCA-5′ (SEQ ID NO: 369) AAT-1499 Target: 5′-GTCTCCCCTCTTCATGGGAAAAGTGGT-3′ (SEQ ID NO: 567) 5′-CUCCCCUCUUCAUGGGAAAAGUGGUGA-3′ (SEQ ID NO: 1162) 3′-GAGGGGAGAAGUACCCUUUUCACCACU-5′ (SEQ ID NO: 370) AAT-1501 Target: 5′-CTCCCCTCTTCATGGGAAAAGTGGTGA-3′ (SEQ ID NO: 568) 5′-UCCCCUCUUCAUGGGAAAAGUGGUGAA-3′ (SEQ ID NO: 1163) 3′-AGGGGAGAAGUACCCUUUUCACCACUU-5′ (SEQ ID NO: 371) AAT-1502 Target: 5′-TCCCCTCTTCATGGGAAAAGTGGTGAA-3′ (SEQ ID NO: 569) 5′-CCCCUCUUCAUGGGAAAAGUGGUGAAU-3′ (SEQ ID NO: 1164) 3′-GGGGAGAAGUACCCUUUUCACCACUUA-5′ (SEQ ID NO: 372) AAT-1503 Target: 5′-CCCCTCTTCATGGGAAAAGTGGTGAAT-3′ (SEQ ID NO: 570) 5′-CCCUCUUCAUGGGAAAAGUGGUGAAUC-3′ (SEQ ID NO: 1165) 3′-GGGAGAAGUACCCUUUUCACCACUUAG-5′ (SEQ ID NO: 373) AAT-1504 Target: 5′-CCCTCTTCATGGGAAAAGTGGTGAATC-3′ (SEQ ID NO: 571) 5′-CCUCUUCAUGGGAAAAGUGGUGAAUCC-3′ (SEQ ID NO: 1166) 3′-GGAGAAGUACCCUUUUCACCACUUAGG-5′ (SEQ ID NO: 374) AAT-1505 Target: 5′-CCTCTTCATGGGAAAAGTGGTGAATCC-3′ (SEQ ID NO: 572) 5′-CUCUUCAUGGGAAAAGUGGUGAAUCCC-3′ (SEQ ID NO: 1167) 3′-GAGAAGUACCCUUUUCACCACUUAGGG-5′ (SEQ ID NO: 375) AAT-1506 Target: 5′-CTCTTCATGGGAAAAGTGGTGAATCCC-3′ (SEQ ID NO: 573) 5′-UCUUCAUGGGAAAAGUGGUGAAUCCCA-3′ (SEQ ID NO: 1168) 3′-AGAAGUACCCUUUUCACCACUUAGGGU-5′ (SEQ ID NO: 376) AAT-1507 Target: 5′-TCTTCATGGGAAAAGTGGTGAATCCCA-3′ (SEQ ID NO: 574) 5′-CUUCAUGGGAAAAGUGGUGAAUCCCAC-3′ (SEQ ID NO: 1169) 3′-GAAGUACCCUUUUCACCACUUAGGGUG-5′ (SEQ ID NO: 377) AAT-1508 Target: 5′-CTTCATGGGAAAAGTGGTGAATCCCAC-3′ (SEQ ID NO: 575) 5′-UUCAUGGGAAAAGUGGUGAAUCCCACC-3′ (SEQ ID NO: 1170) 3′-AAGUACCCUUUUCACCACUUAGGGUGG-5′ (SEQ ID NO: 378) AAT-1509 Target: 5′-TTCATGGGAAAAGTGGTGAATCCCACC-3′ (SEQ ID NO: 576) 5′-UCAUGGGAAAAGUGGUGAAUCCCACCC-3′ (SEQ ID NO: 1171) 3′-AGUACCCUUUUCACCACUUAGGGUGGG-5′ (SEQ ID NO: 379) AAT-1510 Target: 5′-TCATGGGAAAAGTGGTGAATCCCACCC-3′ (SEQ ID NO: 577) 5′-CAUGGGAAAAGUGGUGAAUCCCACCCA-3′ (SEQ ID NO: 1172) 3′-GUACCCUUUUCACCACUUAGGGUGGGU-5′ (SEQ ID NO: 380) AAT-1511 Target: 5′-CATGGGAAAAGTGGTGAATCCCACCCA-3′ (SEQ ID NO: 578) 5′-AUGGGAAAAGUGGUGAAUCCCACCCAA-3′ (SEQ ID NO: 1173) 3′-UACCCUUUUCACCACUUAGGGUGGGUU-5′ (SEQ ID NO: 381) AAT-1512 Target: 5′-ATGGGAAAAGTGGTGAATCCCACCCAA-3′ (SEQ ID NO: 579) 5′-UGGGAAAAGUGGUGAAUCCCACCCAAA-3′ (SEQ ID NO: 1174) 3′-ACCCUUUUCACCACUUAGGGUGGGUUU-5′ (SEQ ID NO: 382) AAT-1513 Target: 5′-TGGGAAAAGTGGTGAATCCCACCCAAA-3′ (SEQ ID NO: 580) 5′-GGGAAAAGUGGUGAAUCCCACCCAAAA-3′ (SEQ ID NO: 1175) 3′-CCCUUUUCACCACUUAGGGUGGGUUUU-5′ (SEQ ID NO: 383) AAT-1514 Target: 5′-GGGAAAAGTGGTGAATCCCACCCAAAA-3′ (SEQ ID NO: 581) 5′-GGAAAAGUGGUGAAUCCCACCCAAAAA-3′ (SEQ ID NO: 1176) 3′-CCUUUUCACCACUUAGGGUGGGUUUUU-5′ (SEQ ID NO: 384) AAT-1515 Target: 5′-GGAAAAGTGGTGAATCCCACCCAAAAA-3′ (SEQ ID NO: 582) 5′-GAAAAGUGGUGAAUCCCACCCAAAAAU-3′ (SEQ ID NO: 1177) 3′-CUUUUCACCACUUAGGGUGGGUUUUUA-5′ (SEQ ID NO: 385) AAT-1516 Target: 5′-GAAAAGTGGTGAATCCCACCCAAAAAT-3′ (SEQ ID NO: 583) 5′-AAAAGUGGUGAAUCCCACCCAAAAAUA-3′ (SEQ ID NO: 1178) 3′-UUUUCACCACUUAGGGUGGGUUUUUAU-5′ (SEQ ID NO: 386) AAT-1517 Target: 5′-AAAAGTGGTGAATCCCACCCAAAAATA-3′ (SEQ ID NO: 584) 5′-UUCGAUAGUUCAAAAUGGUGAAAUUAG-3′ (SEQ ID NO: 1179) 3′-AAGCUAUCAAGUUUUACCACUUUAAUC-5′ (SEQ ID NO: 387) AAT-2872 Target: 5′-TTCGATAGTTCAAAATGGTGAAATTAG-3′ (SEQ ID NO: 585) 5′-UUCAAAAUGGUGAAAUUAGCAAUUCUA-3′ (SEQ ID NO: 1180) 3′-AAGUUUUACCACUUUAAUCGUUAAGAU-5′ (SEQ ID NO: 388) AAT-2880 Target: 5′-TTCAAAATGGTGAAATTAGCAATTCTA-3′ (SEQ ID NO: 586) 5′-AGUUGGUAUGAUGUUCAAGUUAGAUAA-3′ (SEQ ID NO: 1181) 3′-UCAACCAUACUACAAGUUCAAUCUAUU-5′ (SEQ ID NO: 389) AAT-3167 Target: 5′-AGTTGGTATGATGTTCAAGTTAGATAA-3′ (SEQ ID NO: 587) 5′-UUGGUAUGAUGUUCAAGUUAGAUAACA-3′ (SEQ ID NO: 1182) 3′-AACCAUACUACAAGUUCAAUCUAUUGU-5′ (SEQ ID NO: 390) AAT-3169 Target: 5′-TTGGTATGATGTTCAAGTTAGATAACA-3′ (SEQ ID NO: 588) 5′-UGGUAUGAUGUUCAAGUUAGAUAACAA-3′ (SEQ ID NO: 1183) 3′-ACCAUACUACAAGUUCAAUCUAUUGUU-5′ (SEQ ID NO: 391) AAT-3170 Target: 5′-TGGTATGATGTTCAAGTTAGATAACAA-3′ (SEQ ID NO: 589) 5′-GUAUGAUGUUCAAGUUAGAUAACAAAA-3′ (SEQ ID NO: 1184) 3′-CAUACUACAAGUUCAAUCUAUUGUUUU-5′ (SEQ ID NO: 392) AAT-3172 Target: 5′-GTATGATGTTCAAGTTAGATAACAAAA-3′ (SEQ ID NO: 590) 5′-UGAUGUUCAAGUUAGAUAACAAAAUGU-3′ (SEQ ID NO: 1185) 3′-ACUACAAGUUCAAUCUAUUGUUUUACA-5′ (SEQ ID NO: 393) AAT-3175 Target: 5′-TGATGTTCAAGTTAGATAACAAAATGT-3′ (SEQ ID NO: 591) 5′-UUCAAGUUAGAUAACAAAAUGUUUAUA-3′ (SEQ ID NO: 1186) 3′-AAGUUCAAUCUAUUGUUUUACAAAUAU-5′ (SEQ ID NO: 394) AAT-3180 Target: 5′-TTCAAGTTAGATAACAAAATGTTTATA-3′ (SEQ ID NO: 592) 5′-UCAAGUUAGAUAACAAAAUGUUUAUAC-3′ (SEQ ID NO: 1187) 3′-AGUUCAAUCUAUUGUUUUACAAAUAUG-5′ (SEQ ID NO: 395) AAT-3181 Target: 5′-TCAAGTTAGATAACAAAATGTTTATAC-3′ (SEQ ID NO: 593) 5′-CAAGUUAGAUAACAAAAUGUUUAUACC-3′ (SEQ ID NO: 1188) 3′-GUUCAAUCUAUUGUUUUACAAAUAUGG-5′ (SEQ ID NO: 396) AAT-3182 Target: 5′-CAAGTTAGATAACAAAATGTTTATACC-3′ (SEQ ID NO: 594)

TABLE 6 DsiRNA Component 19 Nucleotide Target Sequences in α-1 antitrypsin mRNA AAT-395  19 nt Target #1: 5′-CAUCCCACCAUGAUCAGGA-3′ (SEQ ID NO: 1189) AAT-395  19 nt Target #2: 5′-ACAUCCCACCAUGAUCAGG-3′ (SEQ ID NO: 1387) AAT-395  19 nt Target #3: 5′-UACAUCCCACCAUGAUCAG-3′ (SEQ ID NO: 1585) AAT-475  19 nt Target #1: 5′-CAGCUGGCACACCAGUCCA-3′ (SEQ ID NO: 1190) AAT-475  19 nt Target #2: 5′-CCAGCUGGCACACCAGUCC-3′ (SEQ ID NO: 1388) AAT-475  19 nt Target #3: 5′-GCCAGCUGGCACACCAGUC-3′ (SEQ ID NO: 1586) AAT-477  19 nt Target #1: 5′-GCUGGCACACCAGUCCAAC-3′ (SEQ ID NO: 1191) AAT-477  19 nt Target #2: 5′-AGCUGGCACACCAGUCCAA-3′ (SEQ ID NO: 1389) AAT-477  19 nt Target #3: 5′-CAGCUGGCACACCAGUCCA-3′ (SEQ ID NO: 1587) AAT-480  19 nt Target #1: 5′-GGCACACCAGUCCAACAGC-3′ (SEQ ID NO: 1192) AAT-480  19 nt Target #2: 5′-UGGCACACCAGUCCAACAG-3′ (SEQ ID NO: 1390) AAT-480  19 nt Target #3: 5′-CUGGCACACCAGUCCAACA-3′ (SEQ ID NO: 1588) AAT-481  19 nt Target #1: 5′-GCACACCAGUCCAACAGCA-3′ (SEQ ID NO: 1193) AAT-481  19 nt Target #2: 5′-GGCACACCAGUCCAACAGC-3′ (SEQ ID NO: 1391) AAT-481  19 nt Target #3: 5′-UGGCACACCAGUCCAACAG-3′ (SEQ ID NO: 1589) AAT-482  19 nt Target #1: 5′-CACACCAGUCCAACAGCAC-3′ (SEQ ID NO: 1194) AAT-482  19 nt Target #2: 5′-GCACACCAGUCCAACAGCA-3′ (SEQ ID NO: 1392) AAT-482  19 nt Target #3: 5′-GGCACACCAGUCCAACAGC-3′ (SEQ ID NO: 1590) AAT-483  19 nt Target #1: 5′-ACACCAGUCCAACAGCACC-3′ (SEQ ID NO: 1195) AAT-483  19 nt Target #2: 5′-CACACCAGUCCAACAGCAC-3′ (SEQ ID NO: 1393) AAT-483  19 nt Target #3: 5′-GCACACCAGUCCAACAGCA-3′ (SEQ ID NO: 1591) AAT-484  19 nt Target #1: 5′-CACCAGUCCAACAGCACCA-3′ (SEQ ID NO: 1196) AAT-484  19 nt Target #2: 5′-ACACCAGUCCAACAGCACC-3′ (SEQ ID NO: 1394) AAT-484  19 nt Target #3: 5′-CACACCAGUCCAACAGCAC-3′ (SEQ ID NO: 1592) AAT-500  19 nt Target #1: 5′-CCAAUAUCUUCUUCUCCCC-3′ (SEQ ID NO: 1197) AAT-500  19 nt Target #2: 5′-ACCAAUAUCUUCUUCUCCC-3′ (SEQ ID NO: 1395) AAT-500  19 nt Target #3: 5′-CACCAAUAUCUUCUUCUCC-3′ (SEQ ID NO: 1593) AAT-501  19 nt Target #1: 5′-CAAUAUCUUCUUCUCCCCA-3′ (SEQ ID NO: 1198) AAT-501  19 nt Target #2: 5′-CCAAUAUCUUCUUCUCCCC-3′ (SEQ ID NO: 1396) AAT-501  19 nt Target #3: 5′-ACCAAUAUCUUCUUCUCCC-3′ (SEQ ID NO: 1594) AAT-502  19 nt Target #1: 5′-AAUAUCUUCUUCUCCCCAG-3′ (SEQ ID NO: 1199) AAT-502  19 nt Target #2: 5′-CAAUAUCUUCUUCUCCCCA-3′ (SEQ ID NO: 1397) AAT-502  19 nt Target #3: 5′-CCAAUAUCUUCUUCUCCCC-3′ (SEQ ID NO: 1595) AAT-503  19 nt Target #1: 5′-AUAUCUUCUUCUCCCCAGU-3′ (SEQ ID NO: 1200) AAT-503  19 nt Target #2: 5′-AAUAUCUUCUUCUCCCCAG-3′ (SEQ ID NO: 1398) AAT-503  19 nt Target #3: 5′-CAAUAUCUUCUUCUCCCCA-3′ (SEQ ID NO: 1596) AAT-504  19 nt Target #1: 5′-UAUCUUCUUCUCCCCAGUG-3′ (SEQ ID NO: 1201) AAT-504  19 nt Target #2: 5′-AUAUCUUCUUCUCCCCAGU-3′ (SEQ ID NO: 1399) AAT-504  19 nt Target #3: 5′-AAUAUCUUCUUCUCCCCAG-3′ (SEQ ID NO: 1597) AAT-505  19 nt Target #1: 5′-AUCUUCUUCUCCCCAGUGA-3′ (SEQ ID NO: 1202) AAT-505  19 nt Target #2: 5′-UAUCUUCUUCUCCCCAGUG-3′ (SEQ ID NO: 1400) AAT-505  19 nt Target #3: 5′-AUAUCUUCUUCUCCCCAGU-3′ (SEQ ID NO: 1598) AAT-506  19 nt Target #1: 5′-UCUUCUUCUCCCCAGUGAG-3′ (SEQ ID NO: 1203) AAT-506  19 nt Target #2: 5′-AUCUUCUUCUCCCCAGUGA-3′ (SEQ ID NO: 1401) AAT-506  19 nt Target #3: 5′-UAUCUUCUUCUCCCCAGUG-3′ (SEQ ID NO: 1599) AAT-507  19 nt Target #1: 5′-CUUCUUCUCCCCAGUGAGC-3′ (SEQ ID NO: 1204) AAT-507  19 nt Target #2: 5′-UCUUCUUCUCCCCAGUGAG-3′ (SEQ ID NO: 1402) AAT-507  19 nt Target #3: 5′-AUCUUCUUCUCCCCAGUGA-3′ (SEQ ID NO: 1600) AAT-508  19 nt Target #1: 5′-UUCUUCUCCCCAGUGAGCA-3′ (SEQ ID NO: 1205) AAT-508  19 nt Target #2: 5′-CUUCUUCUCCCCAGUGAGC-3′ (SEQ ID NO: 1403) AAT-508  19 nt Target #3: 5′-UCUUCUUCUCCCCAGUGAG-3′ (SEQ ID NO: 1601) AAT-509  19 nt Target #1: 5′-UCUUCUCCCCAGUGAGCAU-3′ (SEQ ID NO: 1206) AAT-509  19 nt Target #2: 5′-UUCUUCUCCCCAGUGAGCA-3′ (SEQ ID NO: 1404) AAT-509  19 nt Target #3: 5′-CUUCUUCUCCCCAGUGAGC-3′ (SEQ ID NO: 1602) AAT-510  19 nt Target #1: 5′-CUUCUCCCCAGUGAGCAUC-3′ (SEQ ID NO: 1207) AAT-510  19 nt Target #2: 5′-UCUUCUCCCCAGUGAGCAU-3′ (SEQ ID NO: 1405) AAT-510  19 nt Target #3: 5′-UUCUUCUCCCCAGUGAGCA-3′ (SEQ ID NO: 1603) AAT-512  19 nt Target #1: 5′-UCUCCCCAGUGAGCAUCGC-3′ (SEQ ID NO: 1208) AAT-512  19 nt Target #2: 5′-UUCUCCCCAGUGAGCAUCG-3′ (SEQ ID NO: 1406) AAT-512  19 nt Target #3: 5′-CUUCUCCCCAGUGAGCAUC-3′ (SEQ ID NO: 1604) AAT-513  19 nt Target #1: 5′-CUCCCCAGUGAGCAUCGCU-3′ (SEQ ID NO: 1209) AAT-513  19 nt Target #2: 5′-UCUCCCCAGUGAGCAUCGC-3′ (SEQ ID NO: 1407) AAT-513  19 nt Target #3: 5′-UUCUCCCCAGUGAGCAUCG-3′ (SEQ ID NO: 1605) AAT-515  19 nt Target #1: 5′-CCCCAGUGAGCAUCGCUAC-3′ (SEQ ID NO: 1210) AAT-515  19 nt Target #2: 5′-UCCCCAGUGAGCAUCGCUA-3′ (SEQ ID NO: 1408) AAT-515  19 nt Target #3: 5′-CUCCCCAGUGAGCAUCGCU-3′ (SEQ ID NO: 1606) AAT-532  19 nt Target #1: 5′-ACAGCCUUUGCAAUGCUCU-3′ (SEQ ID NO: 1211) AAT-532  19 nt Target #2: 5′-UACAGCCUUUGCAAUGCUC-3′ (SEQ ID NO: 1409) AAT-532  19 nt Target #3: 5′-CUACAGCCUUUGCAAUGCU-3′ (SEQ ID NO: 1607) AAT-540  19 nt Target #1: 5′-UGCAAUGCUCUCCCUGGGG-3′ (SEQ ID NO: 1212) AAT-540  19 nt Target #2: 5′-UUGCAAUGCUCUCCCUGGG-3′ (SEQ ID NO: 1410) AAT-540  19 nt Target #3: 5′-UUUGCAAUGCUCUCCCUGG-3′ (SEQ ID NO: 1608) AAT-581  19 nt Target #1: 5′-AAAUCCUGGAGGGCCUGAA-3′ (SEQ ID NO: 1213) AAT-581  19 nt Target #2: 5′-GAAAUCCUGGAGGGCCUGA-3′ (SEQ ID NO: 1411) AAT-581  19 nt Target #3: 5′-UGAAAUCCUGGAGGGCCUG-3′ (SEQ ID NO: 1609) AAT-582  19 nt Target #1: 5′-AAUCCUGGAGGGCCUGAAU-3′ (SEQ ID NO: 1214) AAT-582  19 nt Target #2: 5′-AAAUCCUGGAGGGCCUGAA-3′ (SEQ ID NO: 1412) AAT-582  19 nt Target #3: 5′-GAAAUCCUGGAGGGCCUGA-3′ (SEQ ID NO: 1610) AAT-583  19 nt Target #1: 5′-AUCCUGGAGGGCCUGAAUU-3′ (SEQ ID NO: 1215) AAT-583  19 nt Target #2: 5′-AAUCCUGGAGGGCCUGAAU-3′ (SEQ ID NO: 1413) AAT-583  19 nt Target #3: 5′-AAAUCCUGGAGGGCCUGAA-3′ (SEQ ID NO: 1611) AAT-585  19 nt Target #1: 5′-CCUGGAGGGCCUGAAUUUC-3′ (SEQ ID NO: 1216) AAT-585  19 nt Target #2: 5′-UCCUGGAGGGCCUGAAUUU-3′ (SEQ ID NO: 1414) AAT-585  19 nt Target #3: 5′-AUCCUGGAGGGCCUGAAUU-3′ (SEQ ID NO: 1612) AAT-586  19 nt Target #1: 5′-CUGGAGGGCCUGAAUUUCA-3′ (SEQ ID NO: 1217) AAT-586  19 nt Target #2: 5′-CCUGGAGGGCCUGAAUUUC-3′ (SEQ ID NO: 1415) AAT-586  19 nt Target #3: 5′-UCCUGGAGGGCCUGAAUUU-3′ (SEQ ID NO: 1613) AAT-587  19 nt Target #1: 5′-UGGAGGGCCUGAAUUUCAA-3′ (SEQ ID NO: 1218) AAT-587  19 nt Target #2: 5′-CUGGAGGGCCUGAAUUUCA-3′ (SEQ ID NO: 1416) AAT-587  19 nt Target #3: 5′-CCUGGAGGGCCUGAAUUUC-3′ (SEQ ID NO: 1614) AAT-634  19 nt Target #1: 5′-CAUGAAGGCUUCCAGGAAC-3′ (SEQ ID NO: 1219) AAT-634  19 nt Target #2: 5′-CCAUGAAGGCUUCCAGGAA-3′ (SEQ ID NO: 1417) AAT-634  19 nt Target #3: 5′-UCCAUGAAGGCUUCCAGGA-3′ (SEQ ID NO: 1615) AAT-637  19 nt Target #1: 5′-GAAGGCUUCCAGGAACUCC-3′ (SEQ ID NO: 1220) AAT-637  19 nt Target #2: 5′-UGAAGGCUUCCAGGAACUC-3′ (SEQ ID NO: 1418) AAT-637  19 nt Target #3: 5′-AUGAAGGCUUCCAGGAACU-3′ (SEQ ID NO: 1616) AAT-638  19 nt Target #1: 5′-AAGGCUUCCAGGAACUCCU-3′ (SEQ ID NO: 1221) AAT-638  19 nt Target #2: 5′-GAAGGCUUCCAGGAACUCC-3′ (SEQ ID NO: 1419) AAT-638  19 nt Target #3: 5′-UGAAGGCUUCCAGGAACUC-3′ (SEQ ID NO: 1617) AAT-671  19 nt Target #1: 5′-AGCCAGACAGCCAGCUCCA-3′ (SEQ ID NO: 1222) AAT-671  19 nt Target #2: 5′-CAGCCAGACAGCCAGCUCC-3′ (SEQ ID NO: 1420) AAT-671  19 nt Target #3: 5′-CCAGCCAGACAGCCAGCUC-3′ (SEQ ID NO: 1618) AAT-673  19 nt Target #1: 5′-CCAGACAGCCAGCUCCAGC-3′ (SEQ ID NO: 1223) AAT-673  19 nt Target #2: 5′-GCCAGACAGCCAGCUCCAG-3′ (SEQ ID NO: 1421) AAT-673  19 nt Target #3: 5′-AGCCAGACAGCCAGCUCCA-3′ (SEQ ID NO: 1619) AAT-675  19 nt Target #1: 5′-AGACAGCCAGCUCCAGCUG-3′ (SEQ ID NO: 1224) AAT-675  19 nt Target #2: 5′-CAGACAGCCAGCUCCAGCU-3′ (SEQ ID NO: 1422) AAT-675  19 nt Target #3: 5′-CCAGACAGCCAGCUCCAGC-3′ (SEQ ID NO: 1620) AAT-676  19 nt Target #1: 5′-GACAGCCAGCUCCAGCUGA-3′ (SEQ ID NO: 1225) AAT-676  19 nt Target #2: 5′-AGACAGCCAGCUCCAGCUG-3′ (SEQ ID NO: 1423) AAT-676  19 nt Target #3: 5′-CAGACAGCCAGCUCCAGCU-3′ (SEQ ID NO: 1621) AAT-734  19 nt Target #1: 5′-UAGUGGAUAAGUUUUUGGA-3′ (SEQ ID NO: 1226) AAT-734  19 nt Target #2: 5′-CUAGUGGAUAAGUUUUUGG-3′ (SEQ ID NO: 1424) AAT-734  19 nt Target #3: 5′-GCUAGUGGAUAAGUUUUUG-3′ (SEQ ID NO: 1622) AAT-735  19 nt Target #1: 5′-AGUGGAUAAGUUUUUGGAG-3′ (SEQ ID NO: 1227) AAT-735  19 nt Target #2: 5′-UAGUGGAUAAGUUUUUGGA-3′ (SEQ ID NO: 1425) AAT-735  19 nt Target #3: 5′-CUAGUGGAUAAGUUUUUGG-3′ (SEQ ID NO: 1623) AAT-736  19 nt Target #1: 5′-GUGGAUAAGUUUUUGGAGG-3′ (SEQ ID NO: 1228) AAT-736  19 nt Target #2: 5′-AGUGGAUAAGUUUUUGGAG-3′ (SEQ ID NO: 1426) AAT-736  19 nt Target #3: 5′-UAGUGGAUAAGUUUUUGGA-3′ (SEQ ID NO: 1624) AAT-737  19 nt Target #1: 5′-UGGAUAAGUUUUUGGAGGA-3′ (SEQ ID NO: 1229) AAT-737  19 nt Target #2: 5′-GUGGAUAAGUUUUUGGAGG-3′ (SEQ ID NO: 1427) AAT-737  19 nt Target #3: 5′-AGUGGAUAAGUUUUUGGAG-3′ (SEQ ID NO: 1625) AAT-738  19 nt Target #1: 5′-GGAUAAGUUUUUGGAGGAU-3′ (SEQ ID NO: 1230) AAT-738  19 nt Target #2: 5′-UGGAUAAGUUUUUGGAGGA-3′ (SEQ ID NO: 1428) AAT-738  19 nt Target #3: 5′-GUGGAUAAGUUUUUGGAGG-3′ (SEQ ID NO: 1626) AAT-739  19 nt Target #1: 5′-GAUAAGUUUUUGGAGGAUG-3′ (SEQ ID NO: 1231) AAT-739  19 nt Target #2: 5′-GGAUAAGUUUUUGGAGGAU-3′ (SEQ ID NO: 1429) AAT-739  19 nt Target #3: 5′-UGGAUAAGUUUUUGGAGGA-3′ (SEQ ID NO: 1627) AAT-740  19 nt Target #1: 5′-AUAAGUUUUUGGAGGAUGU-3′ (SEQ ID NO: 1232) AAT-740  19 nt Target #2: 5′-GAUAAGUUUUUGGAGGAUG-3′ (SEQ ID NO: 1430) AAT-740  19 nt Target #3: 5′-GGAUAAGUUUUUGGAGGAU-3′ (SEQ ID NO: 1628) AAT-767  19 nt Target #1: 5′-UGUACCACUCAGAAGCCUU-3′ (SEQ ID NO: 1233) AAT-767  19 nt Target #2: 5′-UUGUACCACUCAGAAGCCU-3′ (SEQ ID NO: 1431) AAT-767  19 nt Target #3: 5′-GUUGUACCACUCAGAAGCC-3′ (SEQ ID NO: 1629) AAT-768  19 nt Target #1: 5′-GUACCACUCAGAAGCCUUC-3′ (SEQ ID NO: 1234) AAT-768  19 nt Target #2: 5′-UGUACCACUCAGAAGCCUU-3′ (SEQ ID NO: 1432) AAT-768  19 nt Target #3: 5′-UUGUACCACUCAGAAGCCU-3′ (SEQ ID NO: 1630) AAT-850  19 nt Target #1: 5′-ACUCAAGGGAAAAUUGUGG-3′ (SEQ ID NO: 1235) AAT-850  19 nt Target #2: 5′-UACUCAAGGGAAAAUUGUG-3′ (SEQ ID NO: 1433) AAT-850  19 nt Target #3: 5′-GUACUCAAGGGAAAAUUGU-3′ (SEQ ID NO: 1631) AAT-851  19 nt Target #1: 5′-CUCAAGGGAAAAUUGUGGA-3′ (SEQ ID NO: 1236) AAT-851  19 nt Target #2: 5′-ACUCAAGGGAAAAUUGUGG-3′ (SEQ ID NO: 1434) AAT-851  19 nt Target #3: 5′-UACUCAAGGGAAAAUUGUG-3′ (SEQ ID NO: 1632) AAT-852  19 nt Target #1: 5′-UCAAGGGAAAAUUGUGGAU-3′ (SEQ ID NO: 1237) AAT-852  19 nt Target #2: 5′-CUCAAGGGAAAAUUGUGGA-3′ (SEQ ID NO: 1435) AAT-852  19 nt Target #3: 5′-ACUCAAGGGAAAAUUGUGG-3′ (SEQ ID NO: 1633) AAT-853  19 nt Target #1: 5′-CAAGGGAAAAUUGUGGAUU-3′ (SEQ ID NO: 1238) AAT-853  19 nt Target #2: 5′-UCAAGGGAAAAUUGUGGAU-3′ (SEQ ID NO: 1436) AAT-853  19 nt Target #3: 5′-CUCAAGGGAAAAUUGUGGA-3′ (SEQ ID NO: 1634) AAT-854  19 nt Target #1: 5′-AAGGGAAAAUUGUGGAUUU-3′ (SEQ ID NO: 1239) AAT-854  19 nt Target #2: 5′-CAAGGGAAAAUUGUGGAUU-3′ (SEQ ID NO: 1437) AAT-854  19 nt Target #3: 5′-UCAAGGGAAAAUUGUGGAU-3′ (SEQ ID NO: 1635) AAT-855  19 nt Target #1: 5′-AGGGAAAAUUGUGGAUUUG-3′ (SEQ ID NO: 1240) AAT-855  19 nt Target #2: 5′-AAGGGAAAAUUGUGGAUUU-3′ (SEQ ID NO: 1438) AAT-855  19 nt Target #3: 5′-CAAGGGAAAAUUGUGGAUU-3′ (SEQ ID NO: 1636) AAT-856  19 nt Target #1: 5′-GGGAAAAUUGUGGAUUUGG-3′ (SEQ ID NO: 1241) AAT-856  19 nt Target #2: 5′-AGGGAAAAUUGUGGAUUUG-3′ (SEQ ID NO: 1439) AAT-856  19 nt Target #3: 5′-AAGGGAAAAUUGUGGAUUU-3′ (SEQ ID NO: 1637) AAT-857  19 nt Target #1: 5′-GGAAAAUUGUGGAUUUGGU-3′ (SEQ ID NO: 1242) AAT-857  19 nt Target #2: 5′-GGGAAAAUUGUGGAUUUGG-3′ (SEQ ID NO: 1440) AAT-857  19 nt Target #3: 5′-AGGGAAAAUUGUGGAUUUG-3′ (SEQ ID NO: 1638) AAT-858  19 nt Target #1: 5′-GAAAAUUGUGGAUUUGGUC-3′ (SEQ ID NO: 1243) AAT-858  19 nt Target #2: 5′-GGAAAAUUGUGGAUUUGGU-3′ (SEQ ID NO: 1441) AAT-858  19 nt Target #3: 5′-GGGAAAAUUGUGGAUUUGG-3′ (SEQ ID NO: 1639) AAT-859  19 nt Target #1: 5′-AAAAUUGUGGAUUUGGUCA-3′ (SEQ ID NO: 1244) AAT-859  19 nt Target #2: 5′-GAAAAUUGUGGAUUUGGUC-3′ (SEQ ID NO: 1442) AAT-859  19 nt Target #3: 5′-GGAAAAUUGUGGAUUUGGU-3′ (SEQ ID NO: 1640) AAT-860  19 nt Target #1: 5′-AAAUUGUGGAUUUGGUCAA-3′ (SEQ ID NO: 1245) AAT-860  19 nt Target #2: 5′-AAAAUUGUGGAUUUGGUCA-3′ (SEQ ID NO: 1443) AAT-860  19 nt Target #3: 5′-GAAAAUUGUGGAUUUGGUC-3′ (SEQ ID NO: 1641) AAT-861  19 nt Target #1: 5′-AAUUGUGGAUUUGGUCAAG-3′ (SEQ ID NO: 1246) AAT-861  19 nt Target #2: 5′-AAAUUGUGGAUUUGGUCAA-3′ (SEQ ID NO: 1444) AAT-861  19 nt Target #3: 5′-AAAAUUGUGGAUUUGGUCA-3′ (SEQ ID NO: 1642) AAT-862  19 nt Target #1: 5′-AUUGUGGAUUUGGUCAAGG-3′ (SEQ ID NO: 1247) AAT-862  19 nt Target #2: 5′-AAUUGUGGAUUUGGUCAAG-3′ (SEQ ID NO: 1445) AAT-862  19 nt Target #3: 5′-AAAUUGUGGAUUUGGUCAA-3′ (SEQ ID NO: 1643) AAT-863  19 nt Target #1: 5′-UUGUGGAUUUGGUCAAGGA-3′ (SEQ ID NO: 1248) AAT-863  19 nt Target #2: 5′-AUUGUGGAUUUGGUCAAGG-3′ (SEQ ID NO: 1446) AAT-863  19 nt Target #3: 5′-AAUUGUGGAUUUGGUCAAG-3′ (SEQ ID NO: 1644) AAT-864  19 nt Target #1: 5′-UGUGGAUUUGGUCAAGGAG-3′ (SEQ ID NO: 1249) AAT-864  19 nt Target #2: 5′-UUGUGGAUUUGGUCAAGGA-3′ (SEQ ID NO: 1447) AAT-864  19 nt Target #3: 5′-AUUGUGGAUUUGGUCAAGG-3′ (SEQ ID NO: 1645) AAT-865  19 nt Target #1: 5′-GUGGAUUUGGUCAAGGAGC-3′ (SEQ ID NO: 1250) AAT-865  19 nt Target #2: 5′-UGUGGAUUUGGUCAAGGAG-3′ (SEQ ID NO: 1448) AAT-865  19 nt Target #3: 5′-UUGUGGAUUUGGUCAAGGA-3′ (SEQ ID NO: 1646) AAT-866  19 nt Target #1: 5′-UGGAUUUGGUCAAGGAGCU-3′ (SEQ ID NO: 1251) AAT-866  19 nt Target #2: 5′-GUGGAUUUGGUCAAGGAGC-3′ (SEQ ID NO: 1449) AAT-866  19 nt Target #3: 5′-UGUGGAUUUGGUCAAGGAG-3′ (SEQ ID NO: 1647) AAT-867  19 nt Target #1: 5′-GGAUUUGGUCAAGGAGCUU-3′ (SEQ ID NO: 1252) AAT-867  19 nt Target #2: 5′-UGGAUUUGGUCAAGGAGCU-3′ (SEQ ID NO: 1450) AAT-867  19 nt Target #3: 5′-GUGGAUUUGGUCAAGGAGC-3′ (SEQ ID NO: 1648) AAT-868  19 nt Target #1: 5′-GAUUUGGUCAAGGAGCUUG-3′ (SEQ ID NO: 1253) AAT-868  19 nt Target #2: 5′-GGAUUUGGUCAAGGAGCUU-3′ (SEQ ID NO: 1451) AAT-868  19 nt Target #3: 5′-UGGAUUUGGUCAAGGAGCU-3′ (SEQ ID NO: 1649) AAT-869  19 nt Target #1: 5′-AUUUGGUCAAGGAGCUUGA-3′ (SEQ ID NO: 1254) AAT-869  19 nt Target #2: 5′-GAUUUGGUCAAGGAGCUUG-3′ (SEQ ID NO: 1452) AAT-869  19 nt Target #3: 5′-GGAUUUGGUCAAGGAGCUU-3′ (SEQ ID NO: 1650) AAT-870  19 nt Target #1: 5′-UUUGGUCAAGGAGCUUGAC-3′ (SEQ ID NO: 1255) AAT-870  19 nt Target #2: 5′-AUUUGGUCAAGGAGCUUGA-3′ (SEQ ID NO: 1453) AAT-870  19 nt Target #3: 5′-GAUUUGGUCAAGGAGCUUG-3′ (SEQ ID NO: 1651) AAT-871  19 nt Target #1: 5′-UUGGUCAAGGAGCUUGACA-3′ (SEQ ID NO: 1256) AAT-871  19 nt Target #2: 5′-UUUGGUCAAGGAGCUUGAC-3′ (SEQ ID NO: 1454) AAT-871  19 nt Target #3: 5′-AUUUGGUCAAGGAGCUUGA-3′ (SEQ ID NO: 1652) AAT-872  19 nt Target #1: 5′-UGGUCAAGGAGCUUGACAG-3′ (SEQ ID NO: 1257) AAT-872  19 nt Target #2: 5′-UUGGUCAAGGAGCUUGACA-3′ (SEQ ID NO: 1455) AAT-872  19 nt Target #3: 5′-UUUGGUCAAGGAGCUUGAC-3′ (SEQ ID NO: 1653) AAT-896  19 nt Target #1: 5′-CAGUUUUUGCUCUGGUGAA-3′ (SEQ ID NO: 1258) AAT-896  19 nt Target #2: 5′-ACAGUUUUUGCUCUGGUGA-3′ (SEQ ID NO: 1456) AAT-896  19 nt Target #3: 5′-CACAGUUUUUGCUCUGGUG-3′ (SEQ ID NO: 1654) AAT-897  19 nt Target #1: 5′-AGUUUUUGCUCUGGUGAAU-3′ (SEQ ID NO: 1259) AAT-897  19 nt Target #2: 5′-CAGUUUUUGCUCUGGUGAA-3′ (SEQ ID NO: 1457) AAT-897  19 nt Target #3: 5′-ACAGUUUUUGCUCUGGUGA-3′ (SEQ ID NO: 1655) AAT-898  19 nt Target #1: 5′-GUUUUUGCUCUGGUGAAUU-3′ (SEQ ID NO: 1260) AAT-898  19 nt Target #2: 5′-AGUUUUUGCUCUGGUGAAU-3′ (SEQ ID NO: 1458) AAT-898  19 nt Target #3: 5′-CAGUUUUUGCUCUGGUGAA-3′ (SEQ ID NO: 1656) AAT-899  19 nt Target #1: 5′-UUUUUGCUCUGGUGAAUUA-3′ (SEQ ID NO: 1261) AAT-899  19 nt Target #2: 5′-GUUUUUGCUCUGGUGAAUU-3′ (SEQ ID NO: 1459) AAT-899  19 nt Target #3: 5′-AGUUUUUGCUCUGGUGAAU-3′ (SEQ ID NO: 1657) AAT-928  19 nt Target #1: 5′-AAAGGCAAAUGGGAGAGAC-3′ (SEQ ID NO: 1262) AAT-928  19 nt Target #2: 5′-UAAAGGCAAAUGGGAGAGA-3′ (SEQ ID NO: 1460) AAT-928  19 nt Target #3: 5′-UUAAAGGCAAAUGGGAGAG-3′ (SEQ ID NO: 1658) AAT-929  19 nt Target #1: 5′-AAGGCAAAUGGGAGAGACC-3′ (SEQ ID NO: 1263) AAT-929  19 nt Target #2: 5′-AAAGGCAAAUGGGAGAGAC-3′ (SEQ ID NO: 1461) AAT-929  19 nt Target #3: 5′-UAAAGGCAAAUGGGAGAGA-3′ (SEQ ID NO: 1659) AAT-930  19 nt Target #1: 5′-AGGCAAAUGGGAGAGACCC-3′ (SEQ ID NO: 1264) AAT-930  19 nt Target #2: 5′-AAGGCAAAUGGGAGAGACC-3′ (SEQ ID NO: 1462) AAT-930  19 nt Target #3: 5′-AAAGGCAAAUGGGAGAGAC-3′ (SEQ ID NO: 1660) AAT-931  19 nt Target #1: 5′-GGCAAAUGGGAGAGACCCU-3′ (SEQ ID NO: 1265) AAT-931  19 nt Target #2: 5′-AGGCAAAUGGGAGAGACCC-3′ (SEQ ID NO: 1463) AAT-931  19 nt Target #3: 5′-AAGGCAAAUGGGAGAGACC-3′ (SEQ ID NO: 1661) AAT-968  19 nt Target #1: 5′-AGGAAGAGGACUUCCACGU-3′ (SEQ ID NO: 1266) AAT-968  19 nt Target #2: 5′-GAGGAAGAGGACUUCCACG-3′ (SEQ ID NO: 1464) AAT-968  19 nt Target #3: 5′-CGAGGAAGAGGACUUCCAC-3′ (SEQ ID NO: 1662) AAT-969  19 nt Target #1: 5′-GGAAGAGGACUUCCACGUG-3′ (SEQ ID NO: 1267) AAT-969  19 nt Target #2: 5′-AGGAAGAGGACUUCCACGU-3′ (SEQ ID NO: 1465) AAT-969  19 nt Target #3: 5′-GAGGAAGAGGACUUCCACG-3′ (SEQ ID NO: 1663) AAT-970  19 nt Target #1: 5′-GAAGAGGACUUCCACGUGG-3′ (SEQ ID NO: 1268) AAT-970  19 nt Target #2: 5′-GGAAGAGGACUUCCACGUG-3′ (SEQ ID NO: 1466) AAT-970  19 nt Target #3: 5′-AGGAAGAGGACUUCCACGU-3′ (SEQ ID NO: 1664) AAT-971  19 nt Target #1: 5′-AAGAGGACUUCCACGUGGA-3′ (SEQ ID NO: 1269) AAT-971  19 nt Target #2: 5′-GAAGAGGACUUCCACGUGG-3′ (SEQ ID NO: 1467) AAT-971  19 nt Target #3: 5′-GGAAGAGGACUUCCACGUG-3′ (SEQ ID NO: 1665) AAT-973  19 nt Target #1: 5′-GAGGACUUCCACGUGGACC-3′ (SEQ ID NO: 1270) AAT-973  19 nt Target #2: 5′-AGAGGACUUCCACGUGGAC-3′ (SEQ ID NO: 1468) AAT-973  19 nt Target #3: 5′-AAGAGGACUUCCACGUGGA-3′ (SEQ ID NO: 1666) AAT-974  19 nt Target #1: 5′-AGGACUUCCACGUGGACCA-3′ (SEQ ID NO: 1271) AAT-974  19 nt Target #2: 5′-GAGGACUUCCACGUGGACC-3′ (SEQ ID NO: 1469) AAT-974  19 nt Target #3: 5′-AGAGGACUUCCACGUGGAC-3′ (SEQ ID NO: 1667) AAT-976  19 nt Target #1: 5′-GACUUCCACGUGGACCAGG-3′ (SEQ ID NO: 1272) AAT-976  19 nt Target #2: 5′-GGACUUCCACGUGGACCAG-3′ (SEQ ID NO: 1470) AAT-976  19 nt Target #3: 5′-AGGACUUCCACGUGGACCA-3′ (SEQ ID NO: 1668) AAT-1025 19 nt Target #1: 5′-GUUUAGGCAUGUUUAACAU-3′ (SEQ ID NO: 1273) AAT-1025 19 nt Target #2: 5′-CGUUUAGGCAUGUUUAACA-3′ (SEQ ID NO: 1471) AAT-1025 19 nt Target #3: 5′-GCGUUUAGGCAUGUUUAAC-3′ (SEQ ID NO: 1669) AAT-1026 19 nt Target #1: 5′-UUUAGGCAUGUUUAACAUC-3′ (SEQ ID NO: 1274) AAT-1026 19 nt Target #2: 5′-GUUUAGGCAUGUUUAACAU-3′ (SEQ ID NO: 1472) AAT-1026 19 nt Target #3: 5′-CGUUUAGGCAUGUUUAACA-3′ (SEQ ID NO: 1670) AAT-1059 19 nt Target #1: 5′-GCUGUCCAGCUGGGUGCUG-3′ (SEQ ID NO: 1275) AAT-1059 19 nt Target #2: 5′-AGCUGUCCAGCUGGGUGCU-3′ (SEQ ID NO: 1473) AAT-1059 19 nt Target #3: 5′-AAGCUGUCCAGCUGGGUGC-3′ (SEQ ID NO: 1671) AAT-1060 19 nt Target #1: 5′-CUGUCCAGCUGGGUGCUGC-3′ (SEQ ID NO: 1276) AAT-1060 19 nt Target #2: 5′-GCUGUCCAGCUGGGUGCUG-3′ (SEQ ID NO: 1474) AAT-1060 19 nt Target #3: 5′-AGCUGUCCAGCUGGGUGCU-3′ (SEQ ID NO: 1672) AAT-1095 19 nt Target #1: 5′-CAAUGCCACCGCCAUCUUC-3′ (SEQ ID NO: 1277) AAT-1095 19 nt Target #2: 5′-GCAAUGCCACCGCCAUCUU-3′ (SEQ ID NO: 1475) AAT-1095 19 nt Target #3: 5′-GGCAAUGCCACCGCCAUCU-3′ (SEQ ID NO: 1673) AAT-1096 19 nt Target #1: 5′-AAUGCCACCGCCAUCUUCU-3′ (SEQ ID NO: 1278) AAT-1096 19 nt Target #2: 5′-CAAUGCCACCGCCAUCUUC-3′ (SEQ ID NO: 1476) AAT-1096 19 nt Target #3: 5′-GCAAUGCCACCGCCAUCUU-3′ (SEQ ID NO: 1674) AAT-1100 19 nt Target #1: 5′-CCACCGCCAUCUUCUUCCU-3′ (SEQ ID NO: 1279) AAT-1100 19 nt Target #2: 5′-GCCACCGCCAUCUUCUUCC-3′ (SEQ ID NO: 1477) AAT-1100 19 nt Target #3: 5′-UGCCACCGCCAUCUUCUUC-3′ (SEQ ID NO: 1675) AAT-1101 19 nt Target #1: 5′-CACCGCCAUCUUCUUCCUG-3′ (SEQ ID NO: 1280) AAT-1101 19 nt Target #2: 5′-CCACCGCCAUCUUCUUCCU-3′ (SEQ ID NO: 1478) AAT-1101 19 nt Target #3: 5′-GCCACCGCCAUCUUCUUCC-3′ (SEQ ID NO: 1676) AAT-1102 19 nt Target #1: 5′-ACCGCCAUCUUCUUCCUGC-3′ (SEQ ID NO: 1281) AAT-1102 19 nt Target #2: 5′-CACCGCCAUCUUCUUCCUG-3′ (SEQ ID NO: 1479) AAT-1102 19 nt Target #3: 5′-CCACCGCCAUCUUCUUCCU-3′ (SEQ ID NO: 1677) AAT-1103 19 nt Target #1: 5′-CCGCCAUCUUCUUCCUGCC-3′ (SEQ ID NO: 1282) AAT-1103 19 nt Target #2: 5′-ACCGCCAUCUUCUUCCUGC-3′ (SEQ ID NO: 1480) AAT-1103 19 nt Target #3: 5′-CACCGCCAUCUUCUUCCUG-3′ (SEQ ID NO: 1678) AAT-1104 19 nt Target #1: 5′-CGCCAUCUUCUUCCUGCCU-3′ (SEQ ID NO: 1283) AAT-1104 19 nt Target #2: 5′-CCGCCAUCUUCUUCCUGCC-3′ (SEQ ID NO: 1481) AAT-1104 19 nt Target #3: 5′-ACCGCCAUCUUCUUCCUGC-3′ (SEQ ID NO: 1679) AAT-1105 19 nt Target #1: 5′-GCCAUCUUCUUCCUGCCUG-3′ (SEQ ID NO: 1284) AAT-1105 19 nt Target #2: 5′-CGCCAUCUUCUUCCUGCCU-3′ (SEQ ID NO: 1482) AAT-1105 19 nt Target #3: 5′-CCGCCAUCUUCUUCCUGCC-3′ (SEQ ID NO: 1680) AAT-1108 19 nt Target #1: 5′-AUCUUCUUCCUGCCUGAUG-3′ (SEQ ID NO: 1285) AAT-1108 19 nt Target #2: 5′-CAUCUUCUUCCUGCCUGAU-3′ (SEQ ID NO: 1483) AAT-1108 19 nt Target #3: 5′-CCAUCUUCUUCCUGCCUGA-3′ (SEQ ID NO: 1681) AAT-1113 19 nt Target #1: 5′-CUUCCUGCCUGAUGAGGGG-3′ (SEQ ID NO: 1286) AAT-1113 19 nt Target #2: 5′-UCUUCCUGCCUGAUGAGGG-3′ (SEQ ID NO: 1484) AAT-1113 19 nt Target #3: 5′-UUCUUCCUGCCUGAUGAGG-3′ (SEQ ID NO: 1682) AAT-1114 19 nt Target #1: 5′-UUCCUGCCUGAUGAGGGGA-3′ (SEQ ID NO: 1287) AAT-1114 19 nt Target #2: 5′-CUUCCUGCCUGAUGAGGGG-3′ (SEQ ID NO: 1485) AAT-1114 19 nt Target #3: 5′-UCUUCCUGCCUGAUGAGGG-3′ (SEQ ID NO: 1683) AAT-1115 19 nt Target #1: 5′-UCCUGCCUGAUGAGGGGAA-3′ (SEQ ID NO: 1288) AAT-1115 19 nt Target #2: 5′-UUCCUGCCUGAUGAGGGGA-3′ (SEQ ID NO: 1486) AAT-1115 19 nt Target #3: 5′-CUUCCUGCCUGAUGAGGGG-3′ (SEQ ID NO: 1684) AAT-1116 19 nt Target #1: 5′-CCUGCCUGAUGAGGGGAAA-3′ (SEQ ID NO: 1289) AAT-1116 19 nt Target #2: 5′-UCCUGCCUGAUGAGGGGAA-3′ (SEQ ID NO: 1487) AAT-1116 19 nt Target #3: 5′-UUCCUGCCUGAUGAGGGGA-3′ (SEQ ID NO: 1685) AAT-1117 19 nt Target #1: 5′-CUGCCUGAUGAGGGGAAAC-3′ (SEQ ID NO: 1290) AAT-1117 19 nt Target #2: 5′-CCUGCCUGAUGAGGGGAAA-3′ (SEQ ID NO: 1488) AAT-1117 19 nt Target #3: 5′-UCCUGCCUGAUGAGGGGAA-3′ (SEQ ID NO: 1686) AAT-1118 19 nt Target #1: 5′-UGCCUGAUGAGGGGAAACU-3′ (SEQ ID NO: 1291) AAT-1118 19 nt Target #2: 5′-CUGCCUGAUGAGGGGAAAC-3′ (SEQ ID NO: 1489) AAT-1118 19 nt Target #3: 5′-CCUGCCUGAUGAGGGGAAA-3′ (SEQ ID NO: 1687) AAT-1139 19 nt Target #1: 5′-AGCACCUGGAAAAUGAACU-3′ (SEQ ID NO: 1292) AAT-1139 19 nt Target #2: 5′-CAGCACCUGGAAAAUGAAC-3′ (SEQ ID NO: 1490) AAT-1139 19 nt Target #3: 5′-ACAGCACCUGGAAAAUGAA-3′ (SEQ ID NO: 1688) AAT-1140 19 nt Target #1: 5′-GCACCUGGAAAAUGAACUC-3′ (SEQ ID NO: 1293) AAT-1140 19 nt Target #2: 5′-AGCACCUGGAAAAUGAACU-3′ (SEQ ID NO: 1491) AAT-1140 19 nt Target #3: 5′-CAGCACCUGGAAAAUGAAC-3′ (SEQ ID NO: 1689) AAT-1141 19 nt Target #1: 5′-CACCUGGAAAAUGAACUCA-3′ (SEQ ID NO: 1294) AAT-1141 19 nt Target #2: 5′-GCACCUGGAAAAUGAACUC-3′ (SEQ ID NO: 1492) AAT-1141 19 nt Target #3: 5′-AGCACCUGGAAAAUGAACU-3′ (SEQ ID NO: 1690) AAT-1142 19 nt Target #1: 5′-ACCUGGAAAAUGAACUCAC-3′ (SEQ ID NO: 1295) AAT-1142 19 nt Target #2: 5′-CACCUGGAAAAUGAACUCA-3′ (SEQ ID NO: 1493) AAT-1142 19 nt Target #3: 5′-GCACCUGGAAAAUGAACUC-3′ (SEQ ID NO: 1691) AAT-1143 19 nt Target #1: 5′-CCUGGAAAAUGAACUCACC-3′ (SEQ ID NO: 1296) AAT-1143 19 nt Target #2: 5′-ACCUGGAAAAUGAACUCAC-3′ (SEQ ID NO: 1494) AAT-1143 19 nt Target #3: 5′-CACCUGGAAAAUGAACUCA-3′ (SEQ ID NO: 1692) AAT-1166 19 nt Target #1: 5′-AUAUCAUCACCAAGUUCCU-3′ (SEQ ID NO: 1297) AAT-1166 19 nt Target #2: 5′-GAUAUCAUCACCAAGUUCC-3′ (SEQ ID NO: 1495) AAT-1166 19 nt Target #3: 5′-CGAUAUCAUCACCAAGUUC-3′ (SEQ ID NO: 1693) AAT-1167 19 nt Target #1: 5′-UAUCAUCACCAAGUUCCUG-3′ (SEQ ID NO: 1298) AAT-1167 19 nt Target #2: 5′-AUAUCAUCACCAAGUUCCU-3′ (SEQ ID NO: 1496) AAT-1167 19 nt Target #3: 5′-GAUAUCAUCACCAAGUUCC-3′ (SEQ ID NO: 1694) AAT-1168 19 nt Target #1: 5′-AUCAUCACCAAGUUCCUGG-3′ (SEQ ID NO: 1299) AAT-1168 19 nt Target #2: 5′-UAUCAUCACCAAGUUCCUG-3′ (SEQ ID NO: 1497) AAT-1168 19 nt Target #3: 5′-AUAUCAUCACCAAGUUCCU-3′ (SEQ ID NO: 1695) AAT-1169 19 nt Target #1: 5′-UCAUCACCAAGUUCCUGGA-3′ (SEQ ID NO: 1300) AAT-1169 19 nt Target #2: 5′-AUCAUCACCAAGUUCCUGG-3′ (SEQ ID NO: 1498) AAT-1169 19 nt Target #3: 5′-UAUCAUCACCAAGUUCCUG-3′ (SEQ ID NO: 1696) AAT-1170 19 nt Target #1: 5′-CAUCACCAAGUUCCUGGAA-3′ (SEQ ID NO: 1301) AAT-1170 19 nt Target #2: 5′-UCAUCACCAAGUUCCUGGA-3′ (SEQ ID NO: 1499) AAT-1170 19 nt Target #3: 5′-AUCAUCACCAAGUUCCUGG-3′ (SEQ ID NO: 1697) AAT-1171 19 nt Target #1: 5′-AUCACCAAGUUCCUGGAAA-3′ (SEQ ID NO: 1302) AAT-1171 19 nt Target #2: 5′-CAUCACCAAGUUCCUGGAA-3′ (SEQ ID NO: 1500) AAT-1171 19 nt Target #3: 5′-UCAUCACCAAGUUCCUGGA-3′ (SEQ ID NO: 1698) AAT-1172 19 nt Target #1: 5′-UCACCAAGUUCCUGGAAAA-3′ (SEQ ID NO: 1303) AAT-1172 19 nt Target #2: 5′-AUCACCAAGUUCCUGGAAA-3′ (SEQ ID NO: 1501) AAT-1172 19 nt Target #3: 5′-CAUCACCAAGUUCCUGGAA-3′ (SEQ ID NO: 1699) AAT-1173 19 nt Target #1: 5′-CACCAAGUUCCUGGAAAAU-3′ (SEQ ID NO: 1304) AAT-1173 19 nt Target #2: 5′-UCACCAAGUUCCUGGAAAA-3′ (SEQ ID NO: 1502) AAT-1173 19 nt Target #3: 5′-AUCACCAAGUUCCUGGAAA-3′ (SEQ ID NO: 1700) AAT-1174 19 nt Target #1: 5′-ACCAAGUUCCUGGAAAAUG-3′ (SEQ ID NO: 1305) AAT-1174 19 nt Target #2: 5′-CACCAAGUUCCUGGAAAAU-3′ (SEQ ID NO: 1503) AAT-1174 19 nt Target #3: 5′-UCACCAAGUUCCUGGAAAA-3′ (SEQ ID NO: 1701) AAT-1175 19 nt Target #1: 5′-CCAAGUUCCUGGAAAAUGA-3′ (SEQ ID NO: 1306) AAT-1175 19 nt Target #2: 5′-ACCAAGUUCCUGGAAAAUG-3′ (SEQ ID NO: 1504) AAT-1175 19 nt Target #3: 5′-CACCAAGUUCCUGGAAAAU-3′ (SEQ ID NO: 1702) AAT-1286 19 nt Target #1: 5′-AGGUCUUCAGCAAUGGGGC-3′ (SEQ ID NO: 1307) AAT-1286 19 nt Target #2: 5′-AAGGUCUUCAGCAAUGGGG-3′ (SEQ ID NO: 1505) AAT-1286 19 nt Target #3: 5′-UAAGGUCUUCAGCAAUGGG-3′ (SEQ ID NO: 1703) AAT-1296 19 nt Target #1: 5′-CAAUGGGGCUGACCUCUCC-3′ (SEQ ID NO: 1308) AAT-1296 19 nt Target #2: 5′-GCAAUGGGGCUGACCUCUC-3′ (SEQ ID NO: 1506) AAT-1296 19 nt Target #3: 5′-AGCAAUGGGGCUGACCUCU-3′ (SEQ ID NO: 1704) AAT-1297 19 nt Target #1: 5′-AAUGGGGCUGACCUCUCCG-3′ (SEQ ID NO: 1309) AAT-1297 19 nt Target #2: 5′-CAAUGGGGCUGACCUCUCC-3′ (SEQ ID NO: 1507) AAT-1297 19 nt Target #3: 5′-GCAAUGGGGCUGACCUCUC-3′ (SEQ ID NO: 1705) AAT-1298 19 nt Target #1: 5′-AUGGGGCUGACCUCUCCGG-3′ (SEQ ID NO: 1310) AAT-1298 19 nt Target #2: 5′-AAUGGGGCUGACCUCUCCG-3′ (SEQ ID NO: 1508) AAT-1298 19 nt Target #3: 5′-CAAUGGGGCUGACCUCUCC-3′ (SEQ ID NO: 1706) AAT-1324 19 nt Target #1: 5′-GAGGAGGCACCCCUGAAGC-3′ (SEQ ID NO: 1311) AAT-1324 19 nt Target #2: 5′-AGAGGAGGCACCCCUGAAG-3′ (SEQ ID NO: 1509) AAT-1324 19 nt Target #3: 5′-CAGAGGAGGCACCCCUGAA-3′ (SEQ ID NO: 1707) AAT-1326 19 nt Target #1: 5′-GGAGGCACCCCUGAAGCUC-3′ (SEQ ID NO: 1312) AAT-1326 19 nt Target #2: 5′-AGGAGGCACCCCUGAAGCU-3′ (SEQ ID NO: 1510) AAT-1326 19 nt Target #3: 5′-GAGGAGGCACCCCUGAAGC-3′ (SEQ ID NO: 1708) AAT-1336 19 nt Target #1: 5′-CUGAAGCUCUCCAAGGCCG-3′ (SEQ ID NO: 1313) AAT-1336 19 nt Target #2: 5′-CCUGAAGCUCUCCAAGGCC-3′ (SEQ ID NO: 1511) AAT-1336 19 nt Target #3: 5′-CCCUGAAGCUCUCCAAGGC-3′ (SEQ ID NO: 1709) AAT-1353 19 nt Target #1: 5′-CGUGCAUAAGGCUGUGCUG-3′ (SEQ ID NO: 1314) AAT-1353 19 nt Target #2: 5′-CCGUGCAUAAGGCUGUGCU-3′ (SEQ ID NO: 1512) AAT-1353 19 nt Target #3: 5′-GCCGUGCAUAAGGCUGUGC-3′ (SEQ ID NO: 1710) AAT-1354 19 nt Target #1: 5′-GUGCAUAAGGCUGUGCUGA-3′ (SEQ ID NO: 1315) AAT-1354 19 nt Target #2: 5′-CGUGCAUAAGGCUGUGCUG-3′ (SEQ ID NO: 1513) AAT-1354 19 nt Target #3: 5′-CCGUGCAUAAGGCUGUGCU-3′ (SEQ ID NO: 1711) AAT-1355 19 nt Target #1: 5′-UGCAUAAGGCUGUGCUGAC-3′ (SEQ ID NO: 1316) AAT-1355 19 nt Target #2: 5′-GUGCAUAAGGCUGUGCUGA-3′ (SEQ ID NO: 1514) AAT-1355 19 nt Target #3: 5′-CGUGCAUAAGGCUGUGCUG-3′ (SEQ ID NO: 1712) AAT-1356 19 nt Target #1: 5′-GCAUAAGGCUGUGCUGACC-3′ (SEQ ID NO: 1317) AAT-1356 19 nt Target #2: 5′-UGCAUAAGGCUGUGCUGAC-3′ (SEQ ID NO: 1515) AAT-1356 19 nt Target #3: 5′-GUGCAUAAGGCUGUGCUGA-3′ (SEQ ID NO: 1713) AAT-1357 19 nt Target #1: 5′-CAUAAGGCUGUGCUGACCA-3′ (SEQ ID NO: 1318) AAT-1357 19 nt Target #2: 5′-GCAUAAGGCUGUGCUGACC-3′ (SEQ ID NO: 1516) AAT-1357 19 nt Target #3: 5′-UGCAUAAGGCUGUGCUGAC-3′ (SEQ ID NO: 1714) AAT-1358 19 nt Target #1: 5′-AUAAGGCUGUGCUGACCAU-3′ (SEQ ID NO: 1319) AAT-1358 19 nt Target #2: 5′-CAUAAGGCUGUGCUGACCA-3′ (SEQ ID NO: 1517) AAT-1358 19 nt Target #3: 5′-GCAUAAGGCUGUGCUGACC-3′ (SEQ ID NO: 1715) AAT-1359 19 nt Target #1: 5′-UAAGGCUGUGCUGACCAUC-3′ (SEQ ID NO: 1320) AAT-1359 19 nt Target #2: 5′-AUAAGGCUGUGCUGACCAU-3′ (SEQ ID NO: 1518) AAT-1359 19 nt Target #3: 5′-CAUAAGGCUGUGCUGACCA-3′ (SEQ ID NO: 1716) AAT-1360 19 nt Target #1: 5′-AAGGCUGUGCUGACCAUCG-3′ (SEQ ID NO: 1321) AAT-1360 19 nt Target #2: 5′-UAAGGCUGUGCUGACCAUC-3′ (SEQ ID NO: 1519) AAT-1360 19 nt Target #3: 5′-AUAAGGCUGUGCUGACCAU-3′ (SEQ ID NO: 1717) AAT-1361 19 nt Target #1: 5′-AGGCUGUGCUGACCAUCGA-3′ (SEQ ID NO: 1322) AAT-1361 19 nt Target #2: 5′-AAGGCUGUGCUGACCAUCG-3′ (SEQ ID NO: 1520) AAT-1361 19 nt Target #3: 5′-UAAGGCUGUGCUGACCAUC-3′ (SEQ ID NO: 1718) AAT-1390 19 nt Target #1: 5′-ACUGAAGCUGCUGGGGCCA-3′ (SEQ ID NO: 1323) AAT-1390 19 nt Target #2: 5′-GACUGAAGCUGCUGGGGCC-3′ (SEQ ID NO: 1521) AAT-1390 19 nt Target #3: 5′-GGACUGAAGCUGCUGGGGC-3′ (SEQ ID NO: 1719) AAT-1391 19 nt Target #1: 5′-CUGAAGCUGCUGGGGCCAU-3′ (SEQ ID NO: 1324) AAT-1391 19 nt Target #2: 5′-ACUGAAGCUGCUGGGGCCA-3′ (SEQ ID NO: 1522) AAT-1391 19 nt Target #3: 5′-GACUGAAGCUGCUGGGGCC-3′ (SEQ ID NO: 1720) AAT-1392 19 nt Target #1: 5′-UGAAGCUGCUGGGGCCAUG-3′ (SEQ ID NO: 1325) AAT-1392 19 nt Target #2: 5′-CUGAAGCUGCUGGGGCCAU-3′ (SEQ ID NO: 1523) AAT-1392 19 nt Target #3: 5′-ACUGAAGCUGCUGGGGCCA-3′ (SEQ ID NO: 1721) AAT-1393 19 nt Target #1: 5′-GAAGCUGCUGGGGCCAUGU-3′ (SEQ ID NO: 1326) AAT-1393 19 nt Target #2: 5′-UGAAGCUGCUGGGGCCAUG-3′ (SEQ ID NO: 1524) AAT-1393 19 nt Target #3: 5′-CUGAAGCUGCUGGGGCCAU-3′ (SEQ ID NO: 1722) AAT-1394 19 nt Target #1: 5′-AAGCUGCUGGGGCCAUGUU-3′ (SEQ ID NO: 1327) AAT-1394 19 nt Target #2: 5′-GAAGCUGCUGGGGCCAUGU-3′ (SEQ ID NO: 1525) AAT-1394 19 nt Target #3: 5′-UGAAGCUGCUGGGGCCAUG-3′ (SEQ ID NO: 1723) AAT-1395 19 nt Target #1: 5′-AGCUGCUGGGGCCAUGUUU-3′ (SEQ ID NO: 1328) AAT-1395 19 nt Target #2: 5′-AAGCUGCUGGGGCCAUGUU-3′ (SEQ ID NO: 1526) AAT-1395 19 nt Target #3: 5′-GAAGCUGCUGGGGCCAUGU-3′ (SEQ ID NO: 1724) AAT-1405 19 nt Target #1: 5′-GCCAUGUUUUUAGAGGCCA-3′ (SEQ ID NO: 1329) AAT-1405 19 nt Target #2: 5′-GGCCAUGUUUUUAGAGGCC-3′ (SEQ ID NO: 1527) AAT-1405 19 nt Target #3: 5′-GGGCCAUGUUUUUAGAGGC-3′ (SEQ ID NO: 1725) AAT-1406 19 nt Target #1: 5′-CCAUGUUUUUAGAGGCCAU-3′ (SEQ ID NO: 1330) AAT-1406 19 nt Target #2: 5′-GCCAUGUUUUUAGAGGCCA-3′ (SEQ ID NO: 1528) AAT-1406 19 nt Target #3: 5′-GGCCAUGUUUUUAGAGGCC-3′ (SEQ ID NO: 1726) AAT-1407 19 nt Target #1: 5′-CAUGUUUUUAGAGGCCAUA-3′ (SEQ ID NO: 1331) AAT-1407 19 nt Target #2: 5′-CCAUGUUUUUAGAGGCCAU-3′ (SEQ ID NO: 1529) AAT-1407 19 nt Target #3: 5′-GCCAUGUUUUUAGAGGCCA-3′ (SEQ ID NO: 1727) AAT-1408 19 nt Target #1: 5′-AUGUUUUUAGAGGCCAUAC-3′ (SEQ ID NO: 1332) AAT-1408 19 nt Target #2: 5′-CAUGUUUUUAGAGGCCAUA-3′ (SEQ ID NO: 1530) AAT-1408 19 nt Target #3: 5′-CCAUGUUUUUAGAGGCCAU-3′ (SEQ ID NO: 1728) AAT-1409 19 nt Target #1: 5′-UGUUUUUAGAGGCCAUACC-3′ (SEQ ID NO: 1333) AAT-1409 19 nt Target #2: 5′-AUGUUUUUAGAGGCCAUAC-3′ (SEQ ID NO: 1531) AAT-1409 19 nt Target #3: 5′-CAUGUUUUUAGAGGCCAUA-3′ (SEQ ID NO: 1729) AAT-1410 19 nt Target #1: 5′-GUUUUUAGAGGCCAUACCC-3′ (SEQ ID NO: 1334) AAT-1410 19 nt Target #2: 5′-UGUUUUUAGAGGCCAUACC-3′ (SEQ ID NO: 1532) AAT-1410 19 nt Target #3: 5′-AUGUUUUUAGAGGCCAUAC-3′ (SEQ ID NO: 1730) AAT-1411 19 nt Target #1: 5′-UUUUUAGAGGCCAUACCCA-3′ (SEQ ID NO: 1335) AAT-1411 19 nt Target #2: 5′-GUUUUUAGAGGCCAUACCC-3′ (SEQ ID NO: 1533) AAT-1411 19 nt Target #3: 5′-UGUUUUUAGAGGCCAUACC-3′ (SEQ ID NO: 1731) AAT-1412 19 nt Target #1: 5′-UUUUAGAGGCCAUACCCAU-3′ (SEQ ID NO: 1336) AAT-1412 19 nt Target #2: 5′-UUUUUAGAGGCCAUACCCA-3′ (SEQ ID NO: 1534) AAT-1412 19 nt Target #3: 5′-GUUUUUAGAGGCCAUACCC-3′ (SEQ ID NO: 1732) AAT-1413 19 nt Target #1: 5′-UUUAGAGGCCAUACCCAUG-3′ (SEQ ID NO: 1337) AAT-1413 19 nt Target #2: 5′-UUUUAGAGGCCAUACCCAU-3′ (SEQ ID NO: 1535) AAT-1413 19 nt Target #3: 5′-UUUUUAGAGGCCAUACCCA-3′ (SEQ ID NO: 1733) AAT-1414 19 nt Target #1: 5′-UUAGAGGCCAUACCCAUGU-3′ (SEQ ID NO: 1338) AAT-1414 19 nt Target #2: 5′-UUUAGAGGCCAUACCCAUG-3′ (SEQ ID NO: 1536) AAT-1414 19 nt Target #3: 5′-UUUUAGAGGCCAUACCCAU-3′ (SEQ ID NO: 1734) AAT-1415 19 nt Target #1: 5′-UAGAGGCCAUACCCAUGUC-3′ (SEQ ID NO: 1339) AAT-1415 19 nt Target #2: 5′-UUAGAGGCCAUACCCAUGU-3′ (SEQ ID NO: 1537) AAT-1415 19 nt Target #3: 5′-UUUAGAGGCCAUACCCAUG-3′ (SEQ ID NO: 1735) AAT-1416 19 nt Target #1: 5′-AGAGGCCAUACCCAUGUCU-3′ (SEQ ID NO: 1340) AAT-1416 19 nt Target #2: 5′-UAGAGGCCAUACCCAUGUC-3′ (SEQ ID NO: 1538) AAT-1416 19 nt Target #3: 5′-UUAGAGGCCAUACCCAUGU-3′ (SEQ ID NO: 1736) AAT-1452 19 nt Target #1: 5′-GUUCAACAAACCCUUUGUC-3′ (SEQ ID NO: 1341) AAT-1452 19 nt Target #2: 5′-AGUUCAACAAACCCUUUGU-3′ (SEQ ID NO: 1539) AAT-1452 19 nt Target #3: 5′-AAGUUCAACAAACCCUUUG-3′ (SEQ ID NO: 1737) AAT-1453 19 nt Target #1: 5′-UUCAACAAACCCUUUGUCU-3′ (SEQ ID NO: 1342) AAT-1453 19 nt Target #2: 5′-GUUCAACAAACCCUUUGUC-3′ (SEQ ID NO: 1540) AAT-1453 19 nt Target #3: 5′-AGUUCAACAAACCCUUUGU-3′ (SEQ ID NO: 1738) AAT-1454 19 nt Target #1: 5′-UCAACAAACCCUUUGUCUU-3′ (SEQ ID NO: 1343) AAT-1454 19 nt Target #2: 5′-UUCAACAAACCCUUUGUCU-3′ (SEQ ID NO: 1541) AAT-1454 19 nt Target #3: 5′-GUUCAACAAACCCUUUGUC-3′ (SEQ ID NO: 1739) AAT-1455 19 nt Target #1: 5′-CAACAAACCCUUUGUCUUC-3′ (SEQ ID NO: 1344) AAT-1455 19 nt Target #2: 5′-UCAACAAACCCUUUGUCUU-3′ (SEQ ID NO: 1542) AAT-1455 19 nt Target #3: 5′-UUCAACAAACCCUUUGUCU-3′ (SEQ ID NO: 1740) AAT-1456 19 nt Target #1: 5′-AACAAACCCUUUGUCUUCU-3′ (SEQ ID NO: 1345) AAT-1456 19 nt Target #2: 5′-CAACAAACCCUUUGUCUUC-3′ (SEQ ID NO: 1543) AAT-1456 19 nt Target #3: 5′-UCAACAAACCCUUUGUCUU-3′ (SEQ ID NO: 1741) AAT-1457 19 nt Target #1: 5′-ACAAACCCUUUGUCUUCUU-3′ (SEQ ID NO: 1346) AAT-1457 19 nt Target #2: 5′-AACAAACCCUUUGUCUUCU-3′ (SEQ ID NO: 1544) AAT-1457 19 nt Target #3: 5′-CAACAAACCCUUUGUCUUC-3′ (SEQ ID NO: 1742) AAT-1458 19 nt Target #1: 5′-CAAACCCUUUGUCUUCUUA-3′ (SEQ ID NO: 1347) AAT-1458 19 nt Target #2: 5′-ACAAACCCUUUGUCUUCUU-3′ (SEQ ID NO: 1545) AAT-1458 19 nt Target #3: 5′-AACAAACCCUUUGUCUUCU-3′ (SEQ ID NO: 1743) AAT-1459 19 nt Target #1: 5′-AAACCCUUUGUCUUCUUAA-3′ (SEQ ID NO: 1348) AAT-1459 19 nt Target #2: 5′-CAAACCCUUUGUCUUCUUA-3′ (SEQ ID NO: 1546) AAT-1459 19 nt Target #3: 5′-ACAAACCCUUUGUCUUCUU-3′ (SEQ ID NO: 1744) AAT-1460 19 nt Target #1: 5′-AACCCUUUGUCUUCUUAAU-3′ (SEQ ID NO: 1349) AAT-1460 19 nt Target #2: 5′-AAACCCUUUGUCUUCUUAA-3′ (SEQ ID NO: 1547) AAT-1460 19 nt Target #3: 5′-CAAACCCUUUGUCUUCUUA-3′ (SEQ ID NO: 1745) AAT-1489 19 nt Target #1: 5′-AAUACCAAGUCUCCCCUCU-3′ (SEQ ID NO: 1350) AAT-1489 19 nt Target #2: 5′-AAAUACCAAGUCUCCCCUC-3′ (SEQ ID NO: 1548) AAT-1489 19 nt Target #3: 5′-AAAAUACCAAGUCUCCCCU-3′ (SEQ ID NO: 1746) AAT-1490 19 nt Target #1: 5′-AUACCAAGUCUCCCCUCUU-3′ (SEQ ID NO: 1351) AAT-1490 19 nt Target #2: 5′-AAUACCAAGUCUCCCCUCU-3′ (SEQ ID NO: 1549) AAT-1490 19 nt Target #3: 5′-AAAUACCAAGUCUCCCCUC-3′ (SEQ ID NO: 1747) AAT-1491 19 nt Target #1: 5′-UACCAAGUCUCCCCUCUUC-3′ (SEQ ID NO: 1352) AAT-1491 19 nt Target #2: 5′-AUACCAAGUCUCCCCUCUU-3′ (SEQ ID NO: 1550) AAT-1491 19 nt Target #3: 5′-AAUACCAAGUCUCCCCUCU-3′ (SEQ ID NO: 1748) AAT-1492 19 nt Target #1: 5′-ACCAAGUCUCCCCUCUUCA-3′ (SEQ ID NO: 1353) AAT-1492 19 nt Target #2: 5′-UACCAAGUCUCCCCUCUUC-3′ (SEQ ID NO: 1551) AAT-1492 19 nt Target #3: 5′-AUACCAAGUCUCCCCUCUU-3′ (SEQ ID NO: 1749) AAT-1493 19 nt Target #1: 5′-CCAAGUCUCCCCUCUUCAU-3′ (SEQ ID NO: 1354) AAT-1493 19 nt Target #2: 5′-ACCAAGUCUCCCCUCUUCA-3′ (SEQ ID NO: 1552) AAT-1493 19 nt Target #3: 5′-UACCAAGUCUCCCCUCUUC-3′ (SEQ ID NO: 1750) AAT-1494 19 nt Target #1: 5′-CAAGUCUCCCCUCUUCAUG-3′ (SEQ ID NO: 1355) AAT-1494 19 nt Target #2: 5′-CCAAGUCUCCCCUCUUCAU-3′ (SEQ ID NO: 1553) AAT-1494 19 nt Target #3: 5′-ACCAAGUCUCCCCUCUUCA-3′ (SEQ ID NO: 1751) AAT-1495 19 nt Target #1: 5′-AAGUCUCCCCUCUUCAUGG-3′ (SEQ ID NO: 1356) AAT-1495 19 nt Target #2: 5′-CAAGUCUCCCCUCUUCAUG-3′ (SEQ ID NO: 1554) AAT-1495 19 nt Target #3: 5′-CCAAGUCUCCCCUCUUCAU-3′ (SEQ ID NO: 1752) AAT-1496 19 nt Target #1: 5′-AGUCUCCCCUCUUCAUGGG-3′ (SEQ ID NO: 1357) AAT-1496 19 nt Target #2: 5′-AAGUCUCCCCUCUUCAUGG-3′ (SEQ ID NO: 1555) AAT-1496 19 nt Target #3: 5′-CAAGUCUCCCCUCUUCAUG-3′ (SEQ ID NO: 1753) AAT-1497 19 nt Target #1: 5′-GUCUCCCCUCUUCAUGGGA-3′ (SEQ ID NO: 1358) AAT-1497 19 nt Target #2: 5′-AGUCUCCCCUCUUCAUGGG-3′ (SEQ ID NO: 1556) AAT-1497 19 nt Target #3: 5′-AAGUCUCCCCUCUUCAUGG-3′ (SEQ ID NO: 1754) AAT-1499 19 nt Target #1: 5′-CUCCCCUCUUCAUGGGAAA-3′ (SEQ ID NO: 1359) AAT-1499 19 nt Target #2: 5′-UCUCCCCUCUUCAUGGGAA-3′ (SEQ ID NO: 1557) AAT-1499 19 nt Target #3: 5′-GUCUCCCCUCUUCAUGGGA-3′ (SEQ ID NO: 1755) AAT-1501 19 nt Target #1: 5′-CCCCUCUUCAUGGGAAAAG-3′ (SEQ ID NO: 1360) AAT-1501 19 nt Target #2: 5′-UCCCCUCUUCAUGGGAAAA-3′ (SEQ ID NO: 1558) AAT-1501 19 nt Target #3: 5′-CUCCCCUCUUCAUGGGAAA-3′ (SEQ ID NO: 1756) AAT-1502 19 nt Target #1: 5′-CCCUCUUCAUGGGAAAAGU-3′ (SEQ ID NO: 1361) AAT-1502 19 nt Target #2: 5′-CCCCUCUUCAUGGGAAAAG-3′ (SEQ ID NO: 1559) AAT-1502 19 nt Target #3: 5′-UCCCCUCUUCAUGGGAAAA-3′ (SEQ ID NO: 1757) AAT-1503 19 nt Target #1: 5′-CCUCUUCAUGGGAAAAGUG-3′ (SEQ ID NO: 1362) AAT-1503 19 nt Target #2: 5′-CCCUCUUCAUGGGAAAAGU-3′ (SEQ ID NO: 1560) AAT-1503 19 nt Target #3: 5′-CCCCUCUUCAUGGGAAAAG-3′ (SEQ ID NO: 1758) AAT-1504 19 nt Target #1: 5′-CUCUUCAUGGGAAAAGUGG-3′ (SEQ ID NO: 1363) AAT-1504 19 nt Target #2: 5′-CCUCUUCAUGGGAAAAGUG-3′ (SEQ ID NO: 1561) AAT-1504 19 nt Target #3: 5′-CCCUCUUCAUGGGAAAAGU-3′ (SEQ ID NO: 1759) AAT-1505 19 nt Target #1: 5′-UCUUCAUGGGAAAAGUGGU-3′ (SEQ ID NO: 1364) AAT-1505 19 nt Target #2: 5′-CUCUUCAUGGGAAAAGUGG-3′ (SEQ ID NO: 1562) AAT-1505 19 nt Target #3: 5′-CCUCUUCAUGGGAAAAGUG-3′ (SEQ ID NO: 1760) AAT-1506 19 nt Target #1: 5′-CUUCAUGGGAAAAGUGGUG-3′ (SEQ ID NO: 1365) AAT-1506 19 nt Target #2: 5′-UCUUCAUGGGAAAAGUGGU-3′ (SEQ ID NO: 1563) AAT-1506 19 nt Target #3: 5′-CUCUUCAUGGGAAAAGUGG-3′ (SEQ ID NO: 1761) AAT-1507 19 nt Target #1: 5′-UUCAUGGGAAAAGUGGUGA-3′ (SEQ ID NO: 1366) AAT-1507 19 nt Target #2: 5′-CUUCAUGGGAAAAGUGGUG-3′ (SEQ ID NO: 1564) AAT-1507 19 nt Target #3: 5′-UCUUCAUGGGAAAAGUGGU-3′ (SEQ ID NO: 1762) AAT-1508 19 nt Target #1: 5′-UCAUGGGAAAAGUGGUGAA-3′ (SEQ ID NO: 1367) AAT-1508 19 nt Target #2: 5′-UUCAUGGGAAAAGUGGUGA-3′ (SEQ ID NO: 1565) AAT-1508 19 nt Target #3: 5′-CUUCAUGGGAAAAGUGGUG-3′ (SEQ ID NO: 1763) AAT-1509 19 nt Target #1: 5′-CAUGGGAAAAGUGGUGAAU-3′ (SEQ ID NO: 1368) AAT-1509 19 nt Target #2: 5′-UCAUGGGAAAAGUGGUGAA-3′ (SEQ ID NO: 1566) AAT-1509 19 nt Target #3: 5′-UUCAUGGGAAAAGUGGUGA-3′ (SEQ ID NO: 1764) AAT-1510 19 nt Target #1: 5′-AUGGGAAAAGUGGUGAAUC-3′ (SEQ ID NO: 1369) AAT-1510 19 nt Target #2: 5′-CAUGGGAAAAGUGGUGAAU-3′ (SEQ ID NO: 1567) AAT-1510 19 nt Target #3: 5′-UCAUGGGAAAAGUGGUGAA-3′ (SEQ ID NO: 1765) AAT-1511 19 nt Target #1: 5′-UGGGAAAAGUGGUGAAUCC-3′ (SEQ ID NO: 1370) AAT-1511 19 nt Target #2: 5′-AUGGGAAAAGUGGUGAAUC-3′ (SEQ ID NO: 1568) AAT-1511 19 nt Target #3: 5′-CAUGGGAAAAGUGGUGAAU-3′ (SEQ ID NO: 1766) AAT-1512 19 nt Target #1: 5′-GGGAAAAGUGGUGAAUCCC-3′ (SEQ ID NO: 1371) AAT-1512 19 nt Target #2: 5′-UGGGAAAAGUGGUGAAUCC-3′ (SEQ ID NO: 1569) AAT-1512 19 nt Target #3: 5′-AUGGGAAAAGUGGUGAAUC-3′ (SEQ ID NO: 1767) AAT-1513 19 nt Target #1: 5′-GGAAAAGUGGUGAAUCCCA-3′ (SEQ ID NO: 1372) AAT-1513 19 nt Target #2: 5′-GGGAAAAGUGGUGAAUCCC-3′ (SEQ ID NO: 1570) AAT-1513 19 nt Target #3: 5′-UGGGAAAAGUGGUGAAUCC-3′ (SEQ ID NO: 1768) AAT-1514 19 nt Target #1: 5′-GAAAAGUGGUGAAUCCCAC-3′ (SEQ ID NO: 1373) AAT-1514 19 nt Target #2: 5′-GGAAAAGUGGUGAAUCCCA-3′ (SEQ ID NO: 1571) AAT-1514 19 nt Target #3: 5′-GGGAAAAGUGGUGAAUCCC-3′ (SEQ ID NO: 1769) AAT-1515 19 nt Target #1: 5′-AAAAGUGGUGAAUCCCACC-3′ (SEQ ID NO: 1374) AAT-1515 19 nt Target #2: 5′-GAAAAGUGGUGAAUCCCAC-3′ (SEQ ID NO: 1572) AAT-1515 19 nt Target #3: 5′-GGAAAAGUGGUGAAUCCCA-3′ (SEQ ID NO: 1770) AAT-1516 19 nt Target #1: 5′-AAAGUGGUGAAUCCCACCC-3′ (SEQ ID NO: 1375) AAT-1516 19 nt Target #2: 5′-AAAAGUGGUGAAUCCCACC-3′ (SEQ ID NO: 1573) AAT-1516 19 nt Target #3: 5′-GAAAAGUGGUGAAUCCCAC-3′ (SEQ ID NO: 1771) AAT-1517 19 nt Target #1: 5′-AAGUGGUGAAUCCCACCCA-3′ (SEQ ID NO: 1376) AAT-1517 19 nt Target #2: 5′-AAAGUGGUGAAUCCCACCC-3′ (SEQ ID NO: 1574) AAT-1517 19 nt Target #3: 5′-AAAAGUGGUGAAUCCCACC-3′ (SEQ ID NO: 1772) AAT-2872 19 nt Target #1: 5′-CGAUAGUUCAAAAUGGUGA-3′ (SEQ ID NO: 1377) AAT-2872 19 nt Target #2: 5′-UCGAUAGUUCAAAAUGGUG-3′ (SEQ ID NO: 1575) AAT-2872 19 nt Target #3: 5′-UUCGAUAGUUCAAAAUGGU-3′ (SEQ ID NO: 1773) AAT-2880 19 nt Target #1: 5′-CAAAAUGGUGAAAUUAGCA-3′ (SEQ ID NO: 1378) AAT-2880 19 nt Target #2: 5′-UCAAAAUGGUGAAAUUAGC-3′ (SEQ ID NO: 1576) AAT-2880 19 nt Target #3: 5′-UUCAAAAUGGUGAAAUUAG-3′ (SEQ ID NO: 1774) AAT-3167 19 nt Target #1: 5′-UUGGUAUGAUGUUCAAGUU-3′ (SEQ ID NO: 1379) AAT-3167 19 nt Target #2: 5′-GUUGGUAUGAUGUUCAAGU-3′ (SEQ ID NO: 1577) AAT-3167 19 nt Target #3: 5′-AGUUGGUAUGAUGUUCAAG-3′ (SEQ ID NO: 1775) AAT-3169 19 nt Target #1: 5′-GGUAUGAUGUUCAAGUUAG-3′ (SEQ ID NO: 1380) AAT-3169 19 nt Target #2: 5′-UGGUAUGAUGUUCAAGUUA-3′ (SEQ ID NO: 1578) AAT-3169 19 nt Target #3: 5′-UUGGUAUGAUGUUCAAGUU-3′ (SEQ ID NO: 1776) AAT-3170 19 nt Target #1: 5′-GUAUGAUGUUCAAGUUAGA-3′ (SEQ ID NO: 1381) AAT-3170 19 nt Target #2: 5′-GGUAUGAUGUUCAAGUUAG-3′ (SEQ ID NO: 1579) AAT-3170 19 nt Target #3: 5′-UGGUAUGAUGUUCAAGUUA-3′ (SEQ ID NO: 1777) AAT-3172 19 nt Target #1: 5′-AUGAUGUUCAAGUUAGAUA-3′ (SEQ ID NO: 1382) AAT-3172 19 nt Target #2: 5′-UAUGAUGUUCAAGUUAGAU-3′ (SEQ ID NO: 1580) AAT-3172 19 nt Target #3: 5′-GUAUGAUGUUCAAGUUAGA-3′ (SEQ ID NO: 1778) AAT-3175 19 nt Target #1: 5′-AUGUUCAAGUUAGAUAACA-3′ (SEQ ID NO: 1383) AAT-3175 19 nt Target #2: 5′-GAUGUUCAAGUUAGAUAAC-3′ (SEQ ID NO: 1581) AAT-3175 19 nt Target #3: 5′-UGAUGUUCAAGUUAGAUAA-3′ (SEQ ID NO: 1779) AAT-3180 19 nt Target #1: 5′-CAAGUUAGAUAACAAAAUG-3′ (SEQ ID NO: 1384) AAT-3180 19 nt Target #2: 5′-UCAAGUUAGAUAACAAAAU-3′ (SEQ ID NO: 1582) AAT-3180 19 nt Target #3: 5′-UUCAAGUUAGAUAACAAAA-3′ (SEQ ID NO: 1780) AAT-3181 19 nt Target #1: 5′-AAGUUAGAUAACAAAAUGU-3′ (SEQ ID NO: 1385) AAT-3181 19 nt Target #2: 5′-CAAGUUAGAUAACAAAAUG-3′ (SEQ ID NO: 1583) AAT-3181 19 nt Target #3: 5′-UCAAGUUAGAUAACAAAAU-3′ (SEQ ID NO: 1781) AAT-3182 19 nt Target #1: 5′-AGUUAGAUAACAAAAUGUU-3′ (SEQ ID NO: 1386) AAT-3182 19 nt Target #2: 5′-AAGUUAGAUAACAAAAUGU-3′ (SEQ ID NO: 1584) AAT-3182 19 nt Target #3: 5′-CAAGUUAGAUAACAAAAUG-3′ (SEQ ID NO: 1782)

TABLE 7  Additional Human Anti-α-1 antitrypsin DsiRNA Agents (Asymmetrics)  5′-GCGUUUAGGCAUGUUUAACAUCCag-3′ (SEQ ID NO: 1783) 3′-UUCGCAAAUCCGUACAAAUUGUAGGUC-5′ (SEQ ID NO: 1814) AAT-1023 Target: 5′-AAGCGTTTAGGCATGTTTAACATCCAG-3′ (SEQ ID NO: 1845) 5′-CGUUUAGGCAUGUUUAACAUCCAgc-3′ (SEQ ID NO: 1784) 3′-UCGCAAAUCCGUACAAAUUGUAGGUCG-5′ (SEQ ID NO: 1815) AAT-1024 Target: 5′-AGCGTTTAGGCATGTTTAACATCCAGC-3′ (SEQ ID NO: 1846) 5′-GGCCAUGUUUUUAGAGGCCAUACcc-3′ (SEQ ID NO: 1785) 3′-CCCCGGUACAAAAAUCUCCGGUAUGGG-5′ (SEQ ID NO: 1816) AAT-1404 Target: 5′-GGGGCCATGTTTTTAGAGGCCATACCC-3′ (SEQ ID NO: 1847) 5′-ACCCUUUGUCUUCUUAAUGAUUGaa-3′ (SEQ ID NO: 1786) 3′-UUUGGGAAACAGAAGAAUUACUAACUU-5′ (SEQ ID NO: 1817) AAT-1461 Target: 5′-AAACCCTTTGTCTTCTTAATGATTGAA-3′ (SEQ ID NO: 1848) 5′-CCCUUUGUCUUCUUAAUGAUUGAac-3′ (SEQ ID NO: 1787) 3′-UUGGGAAACAGAAGAAUUACUAACUUG-5′ (SEQ ID NO: 1818) AAT-1462 Target: 5′-AACCCTTTGTCTTCTTAATGATTGAAC-3′ (SEQ ID NO: 1849) 5′-CCUUUGUCUUCUUAAUGAUUGAAca-3′ (SEQ ID NO: 1788) 3′-UGGGAAACAGAAGAAUUACUAACUUGU-5′ (SEQ ID NO: 1819) AAT-1463 Target: 5′-ACCCTTTGTCTTCTTAATGATTGAACA-3′ (SEQ ID NO: 1850) 5′-CUUUGUCUUCUUAAUGAUUGAACaa-3′ (SEQ ID NO: 1789) 3′-GGGAAACAGAAGAAUUACUAACUUGUU-5′ (SEQ ID NO: 1820) AAT-1464 Target: 5′-CCCTTTGTCTTCTTAATGATTGAACAA-3′ (SEQ ID NO: 1851) 5′-UUUGUCUUCUUAAUGAUUGAACAaa-3′ (SEQ ID NO: 1790) 3′-GGAAACAGAAGAAUUACUAACUUGUUU-5′ (SEQ ID NO: 1821) AAT-1465 Target: 5′-CCTTTGTCTTCTTAATGATTGAACAAA-3′ (SEQ ID NO: 1852) 5′-UUGUCUUCUUAAUGAUUGAACAAaa-3′ (SEQ ID NO: 1791) 3′-GAAACAGAAGAAUUACUAACUUGUUUU-5′ (SEQ ID NO: 1822) AAT-1466 Target: 5′-CTTTGTCTTCTTAATGATTGAACAAAA-3′ (SEQ ID NO: 1853) 5′-UGUCUUCUUAAUGAUUGAACAAAat-3′ (SEQ ID NO: 1792) 3′-AAACAGAAGAAUUACUAACUUGUUUUA-5′ (SEQ ID NO: 1823) AAT-1467 Target: 5′-TTTGTCTTCTTAATGATTGAACAAAAT-3′ (SEQ ID NO: 1854) 5′-GUCUUCUUAAUGAUUGAACAAAAta-3′ (SEQ ID NO: 1793) 3′-AACAGAAGAAUUACUAACUUGUUUUAU-5′ (SEQ ID NO: 1824) AAT-1468 Target: 5′-TTGTCTTCTTAATGATTGAACAAAATA-3′ (SEQ ID NO: 1855) 5′-UCUUCUUAAUGAUUGAACAAAAUac-3′ (SEQ ID NO: 1794) 3′-ACAGAAGAAUUACUAACUUGUUUUAUG-5′ (SEQ ID NO: 1825) AAT-1469 Target: 5′-TGTCTTCTTAATGATTGAACAAAATAC-3′ (SEQ ID NO: 1856) 5′-CUUCUUAAUGAUUGAACAAAAUAcc-3′ (SEQ ID NO: 1795) 3′-CAGAAGAAUUACUAACUUGUUUUAUGG-5′ (SEQ ID NO: 1826) AAT-1470 Target: 5′-GTCTTCTTAATGATTGAACAAAATACC-3′ (SEQ ID NO: 1857) 5′-UUCUUAAUGAUUGAACAAAAUACca-3′ (SEQ ID NO: 1796) 3′-AGAAGAAUUACUAACUUGUUUUAUGGU-5′ (SEQ ID NO: 1827) AAT-1471 Target: 5′-TCTTCTTAATGATTGAACAAAATACCA-3′ (SEQ ID NO: 1858) 5′-UCUUAAUGAUUGAACAAAAUACCaa-3′ (SEQ ID NO: 1797) 3′-GAAGAAUUACUAACUUGUUUUAUGGUU-5′ (SEQ ID NO: 1828) AAT-1472 Target: 5′-CTTCTTAATGATTGAACAAAATACCAA-3′ (SEQ ID NO: 1859) 5′-CUUAAUGAUUGAACAAAAUACCAag-3′ (SEQ ID NO: 1798) 3′-AAGAAUUACUAACUUGUUUUAUGGUUC-5′ (SEQ ID NO: 1829) AAT-1473 Target: 5′-TTCTTAATGATTGAACAAAATACCAAG-3′ (SEQ ID NO: 1860) 5′-UUAAUGAUUGAACAAAAUACCAAgt-3′ (SEQ ID NO: 1799) 3′-AGAAUUACUAACUUGUUUUAUGGUUCA-5′ (SEQ ID NO: 1830) AAT-1474 Target: 5′-TCTTAATGATTGAACAAAATACCAAGT-3′ (SEQ ID NO: 1861) 5′-UAAUGAUUGAACAAAAUACCAAGtc-3′ (SEQ ID NO: 1800) 3′-GAAUUACUAACUUGUUUUAUGGUUCAG-5′ (SEQ ID NO: 1831) AAT-1475 Target: 5′-CTTAATGATTGAACAAAATACCAAGTC-3′ (SEQ ID NO: 1862) 5′-AAUGAUUGAACAAAAUACCAAGUct-3′ (SEQ ID NO: 1801) 3′-AAUUACUAACUUGUUUUAUGGUUCAGA-5′ (SEQ ID NO: 1832) AAT-1476 Target: 5′-TTAATGATTGAACAAAATACCAAGTCT-3′ (SEQ ID NO: 1863) 5′-AUGAUUGAACAAAAUACCAAGUCtc-3′ (SEQ ID NO: 1802) 3′-AUUACUAACUUGUUUUAUGGUUCAGAG-5′ (SEQ ID NO: 1833) AAT-1477 Target: 5′-TAATGATTGAACAAAATACCAAGTCTC-3′ (SEQ ID NO: 1864) 5′-UGAUUGAACAAAAUACCAAGUCUcc-3′ (SEQ ID NO: 1803) 3′-UUACUAACUUGUUUUAUGGUUCAGAGG-5′ (SEQ ID NO: 1834) AAT-1478 Target: 5′-AATGATTGAACAAAATACCAAGTCTCC-3′ (SEQ ID NO: 1865) 5′-GAUUGAACAAAAUACCAAGUCUCcc-3′ (SEQ ID NO: 1804) 3′-UACUAACUUGUUUUAUGGUUCAGAGGG-5′ (SEQ ID NO: 1835) AAT-1479 Target: 5′-ATGATTGAACAAAATACCAAGTCTCCC-3′ (SEQ ID NO: 1866) 5′-AUUGAACAAAAUACCAAGUCUCCcc-3′ (SEQ ID NO: 1805) 3′-ACUAACUUGUUUUAUGGUUCAGAGGGG-5′ (SEQ ID NO: 1836) AAT-1480 Target: 5′-TGATTGAACAAAATACCAAGTCTCCCC-3′ (SEQ ID NO: 1867) 5′-UUGAACAAAAUACCAAGUCUCCCct-3′ (SEQ ID NO: 1806) 3′-CUAACUUGUUUUAUGGUUCAGAGGGGA-5′ (SEQ ID NO: 1837) AAT-1481 Target: 5′-GATTGAACAAAATACCAAGTCTCCCCT-3′ (SEQ ID NO: 1868) 5′-UGAACAAAAUACCAAGUCUCCCCtc-3′ (SEQ ID NO: 1807) 3′-UAACUUGUUUUAUGGUUCAGAGGGGAG-5′ (SEQ ID NO: 1838) AAT-1482 Target: 5′-ATTGAACAAAATACCAAGTCTCCCCTC-3′ (SEQ ID NO: 1869) 5′-GAACAAAAUACCAAGUCUCCCCUct-3′ (SEQ ID NO: 1808) 3′-AACUUGUUUUAUGGUUCAGAGGGGAGA-5′ (SEQ ID NO: 1839) AAT-1483 Target: 5′-TTGAACAAAATACCAAGTCTCCCCTCT-3′ (SEQ ID NO: 1870) 5′-AACAAAAUACCAAGUCUCCCCUCtt-3′ (SEQ ID NO: 1809) 3′-ACUUGUUUUAUGGUUCAGAGGGGAGAA-5′ (SEQ ID NO: 1840) AAT-1484 Target: 5′-TGAACAAAATACCAAGTCTCCCCTCTT-3′ (SEQ ID NO: 1871) 5′-ACAAAAUACCAAGUCUCCCCUCUtc-3′ (SEQ ID NO: 1810) 3′-CUUGUUUUAUGGUUCAGAGGGGAGAAG-5′ (SEQ ID NO: 1841) AAT-1485 Target: 5′-GAACAAAATACCAAGTCTCCCCTCTTC-3′ (SEQ ID NO: 1872) 5′-CAAAAUACCAAGUCUCCCCUCUUca-3′ (SEQ ID NO: 1811) 3′-UUGUUUUAUGGUUCAGAGGGGAGAAGU-5′ (SEQ ID NO: 1842) AAT-1486 Target: 5′-AACAAAATACCAAGTCTCCCCTCTTCA-3′ (SEQ ID NO: 1873) 5′-AAAAUACCAAGUCUCCCCUCUUCat-3′ (SEQ ID NO: 1812) 3′-UGUUUUAUGGUUCAGAGGGGAGAAGUA-5′ (SEQ ID NO: 1843) AAT-1487 Target: 5′-ACAAAATACCAAGTCTCCCCTCTTCAT-3′ (SEQ ID NO: 1874) 5′-AAAUACCAAGUCUCCCCUCUUCAtg-3′ (SEQ ID NO: 1813) 3′-GUUUUAUGGUUCAGAGGGGAGAAGUAC-5′ (SEQ ID NO: 1844) AAT-1488 Target: 5′-CAAAATACCAAGTCTCCCCTCTTCATG-3′ (SEQ ID NO: 1875) 

TABLE 8  Additional Human Anti-α-1 antitrypsin DsiRNAs,  Unmodified Duplexes (Asymmetrics)  5′-GCGUUUAGGCAUGUUUAACAUCCAG-3′ (SEQ ID NO: 1876) 3′-UUCGCAAAUCCGUACAAAUUGUAGGUC-5′ (SEQ ID NO: 1814) AAT-1023 Target: 5′-AAGCGTTTAGGCATGTTTAACATCCAG-3′ (SEQ ID NO: 1845) 5′-CGUUUAGGCAUGUUUAACAUCCAGC-3′ (SEQ ID NO: 1877) 3′-UCGCAAAUCCGUACAAAUUGUAGGUCG-5′ (SEQ ID NO: 1815) AAT-1024 Target: 5′-AGCGTTTAGGCATGTTTAACATCCAGC-3′ (SEQ ID NO: 1846) 5′-GGCCAUGUUUUUAGAGGCCAUACCC-3′ (SEQ ID NO: 1878) 3′-CCCCGGUACAAAAAUCUCCGGUAUGGG-5′ (SEQ ID NO: 1816) AAT-1404 Target: 5′-GGGGCCATGTTTTTAGAGGCCATACCC-3′ (SEQ ID NO: 1847) 5′-ACCCUUUGUCUUCUUAAUGAUUGAA-3′ (SEQ ID NO: 1879) 3′-UUUGGGAAACAGAAGAAUUACUAACUU-5′ (SEQ ID NO: 1817) AAT-1461 Target: 5′-AAACCCTTTGTCTTCTTAATGATTGAA-3′ (SEQ ID NO: 1848) 5′-CCCUUUGUCUUCUUAAUGAUUGAAC-3′ (SEQ ID NO: 1880) 3′-UUGGGAAACAGAAGAAUUACUAACUUG-5′ (SEQ ID NO: 1818) AAT-1462 Target: 5′-AACCCTTTGTCTTCTTAATGATTGAAC-3′ (SEQ ID NO: 1849) 5′-CCUUUGUCUUCUUAAUGAUUGAACA-3′(SEQ ID NO: 1881) 3′-UGGGAAACAGAAGAAUUACUAACUUGU-5′ (SEQ ID NO: 1819) AAT-1463 Target: 5′-ACCCTTTGTCTTCTTAATGATTGAACA-3′ (SEQ ID NO: 1850) 5′-CUUUGUCUUCUUAAUGAUUGAACAA-3′ (SEQ ID NO: 1882) 3′-GGGAAACAGAAGAAUUACUAACUUGUU-5′(SEQ ID NO: 1820) AAT-1464 Target: 5′-CCCTTTGTCTTCTTAATGATTGAACAA-3′ (SEQ ID NO: 1851) 5′-UUUGUCUUCUUAAUGAUUGAACAAA-3′ (SEQ ID NO: 1883) 3′-GGAAACAGAAGAAUUACUAACUUGUUU-5′(SEQ ID NO: 1821) AAT-1465 Target: 5′-CCTTTGTCTTCTTAATGATTGAACAAA-3′ (SEQ ID NO: 1852) 5′-UUGUCUUCUUAAUGAUUGAACAAAA-3′ (SEQ ID NO: 1884) 3′-GAAACAGAAGAAUUACUAACUUGUUUU-5′ (SEQ ID NO: 1822) AAT-1466 Target: 5′-CTTTGTCTTCTTAATGATTGAACAAAA-3′ (SEQ ID NO: 1853) 5′-UGUCUUCUUAAUGAUUGAACAAAAU-3′ (SEQ ID NO: 1885) 3′-AAACAGAAGAAUUACUAACUUGUUUUA-5′ (SEQ ID NO: 1823) AAT-1467 Target: 5′-TTTGTCTTCTTAATGATTGAACAAAAT-3′ (SEQ ID NO: 1854) 5′-GUCUUCUUAAUGAUUGAACAAAAUA-3′(SEQ ID NO: 1886) 3′-AACAGAAGAAUUACUAACUUGUUUUAU-5′ (SEQ ID NO: 1824) AAT-1468 Target: 5′-TTGTCTTCTTAATGATTGAACAAAATA-3′ (SEQ ID NO: 1855) 5′-UCUUCUUAAUGAUUGAACAAAAUAC-3′ (SEQ ID NO: 1887) 3′-ACAGAAGAAUUACUAACUUGUUUUAUG-5′ (SEQ ID NO: 1825) AAT-1469 Target: 5′-TGTCTTCTTAATGATTGAACAAAATAC-3′ (SEQ ID NO: 1856) 5′-CUUCUUAAUGAUUGAACAAAAUACC-3′ (SEQ ID NO: 1888) 3′-CAGAAGAAUUACUAACUUGUUUUAUGG-5′ (SEQ ID NO: 1826) AAT-1470 Target: 5′-GTCTTCTTAATGATTGAACAAAATACC-3′ (SEQ ID NO: 1857) 5′-UUCUUAAUGAUUGAACAAAAUACCA-3′ (SEQ ID NO: 1889) 3′-AGAAGAAUUACUAACUUGUUUUAUGGU-5′ (SEQ ID NO: 1827) AAT-1471 Target: 5′-TCTTCTTAATGATTGAACAAAATACCA-3′ (SEQ ID NO: 1858) 5′-UCUUAAUGAUUGAACAAAAUACCAA-3′ (SEQ ID NO: 1890) 3′-GAAGAAUUACUAACUUGUUUUAUGGUU-5′ (SEQ ID NO: 1828) AAT-1472 Target: 5′-CTTCTTAATGATTGAACAAAATACCAA-3′ (SEQ ID NO: 1859) 5′-CUUAAUGAUUGAACAAAAUACCAAG-3′ (SEQ ID NO: 1891) 3′-AAGAAUUACUAACUUGUUUUAUGGUUC-5′ (SEQ ID NO: 1829) AAT-1473 Target: 5′-TTCTTAATGATTGAACAAAATACCAAG-3′ (SEQ ID NO: 1860) 5′-UUAAUGAUUGAACAAAAUACCAAGU-3′ (SEQ ID NO: 1892) 3′-AGAAUUACUAACUUGUUUUAUGGUUCA-5′ (SEQ ID NO: 1830) AAT-1474 Target: 5′-TCTTAATGATTGAACAAAATACCAAGT-3′ (SEQ ID NO: 1861) 5′-UAAUGAUUGAACAAAAUACCAAGUC-3′ (SEQ ID NO: 1893) 3′-GAAUUACUAACUUGUUUUAUGGUUCAG-5′ (SEQ ID NO: 1831) AAT-1475 Target: 5′-CTTAATGATTGAACAAAATACCAAGTC-3′ (SEQ ID NO: 1862) 5′-AAUGAUUGAACAAAAUACCAAGUCU-3′ (SEQ ID NO: 1894) 3′-AAUUACUAACUUGUUUUAUGGUUCAGA-5′ (SEQ ID NO: 1832) AAT-1476 Target: 5′-TTAATGATTGAACAAAATACCAAGTCT-3′ (SEQ ID NO: 1863) 5′-AUGAUUGAACAAAAUACCAAGUCUC-3′ (SEQ ID NO: 1895) 3′-AUUACUAACUUGUUUUAUGGUUCAGAG-5′ (SEQ ID NO: 1833) AAT-1477 Target: 5′-TAATGATTGAACAAAATACCAAGTCTC-3′ (SEQ ID NO: 1864) 5′-UGAUUGAACAAAAUACCAAGUCUCC-3′ (SEQ ID NO: 1896) 3′-UUACUAACUUGUUUUAUGGUUCAGAGG-5′ (SEQ ID NO: 1834) AAT-1478 Target: 5′-AATGATTGAACAAAATACCAAGTCTCC-3′ (SEQ ID NO: 1865) 5′-GAUUGAACAAAAUACCAAGUCUCCC-3′ (SEQ ID NO: 1897) 3′-UACUAACUUGUUUUAUGGUUCAGAGGG-5′ (SEQ ID NO: 1835) AAT-1479 Target: 5′-ATGATTGAACAAAATACCAAGTCTCCC-3′ (SEQ ID NO: 1866) 5′-AUUGAACAAAAUACCAAGUCUCCCC-3′ (SEQ ID NO: 1898) 3′-ACUAACUUGUUUUAUGGUUCAGAGGGG-5′ (SEQ ID NO: 1836) AAT-1480 Target: 5′-TGATTGAACAAAATACCAAGTCTCCCC-3′ (SEQ ID NO: 1867) 5′-UUGAACAAAAUACCAAGUCUCCCCU-3′ (SEQ ID NO: 1899) 3′-CUAACUUGUUUUAUGGUUCAGAGGGGA-5′ (SEQ ID NO: 1837) AAT-1481 Target: 5′-GATTGAACAAAATACCAAGTCTCCCCT-3′ (SEQ ID NO: 1868) 5′-UGAACAAAAUACCAAGUCUCCCCUC-3′ (SEQ ID NO: 1900) 3′-UAACUUGUUUUAUGGUUCAGAGGGGAG-5′ (SEQ ID NO: 1838) AAT-1482 Target: 5′-ATTGAACAAAATACCAAGTCTCCCCTC-3′ (SEQ ID NO: 1869) 5′-GAACAAAAUACCAAGUCUCCCCUCU-3′ (SEQ ID NO: 1901) 3′-AACUUGUUUUAUGGUUCAGAGGGGAGA-5′ (SEQ ID NO: 1839) AAT-1483 Target: 5′-TTGAACAAAATACCAAGTCTCCCCTCT-3′ (SEQ ID NO: 1870) 5′-AACAAAAUACCAAGUCUCCCCUCUU-3′ (SEQ ID NO: 1902) 3′-ACUUGUUUUAUGGUUCAGAGGGGAGAA-5′ (SEQ ID NO: 1840) AAT-1484 Target: 5′-TGAACAAAATACCAAGTCTCCCCTCTT-3′ (SEQ ID NO: 1871) 5′-ACAAAAUACCAAGUCUCCCCUCUUC-3′ (SEQ ID NO: 1903) 3′-CUUGUUUUAUGGUUCAGAGGGGAGAAG-5′ (SEQ ID NO: 1841) AAT-1485 Target: 5′-GAACAAAATACCAAGTCTCCCCTCTTC-3′ (SEQ ID NO: 1872) 5′-CAAAAUACCAAGUCUCCCCUCUUCA-3′ (SEQ ID NO: 1904) 3′-UUGUUUUAUGGUUCAGAGGGGAGAAGU-5′ (SEQ ID NO: 1842) AAT-1486 Target: 5′-AACAAAATACCAAGTCTCCCCTCTTCA-3′ (SEQ ID NO: 1873) 5′-AAAAUACCAAGUCUCCCCUCUUCAU-3′ (SEQ ID NO: 1905) 3′-UGUUUUAUGGUUCAGAGGGGAGAAGUA-5′ (SEQ ID NO: 1843) AAT-1487 Target: 5′-ACAAAATACCAAGTCTCCCCTCTTCAT-3′ (SEQ ID NO: 1874) 5′-AAAUACCAAGUCUCCCCUCUUCAUG-3′ (SEQ ID NO: 1906) 3′-GUUUUAUGGUUCAGAGGGGAGAAGUAC-5′ (SEQ ID NO: 1844) AAT-1488 Target: 5′-CAAAATACCAAGTCTCCCCTCTTCATG-3′ (SEQ ID NO: 1875) 

TABLE 9  Additional DsiRNA Target Sequences (21mers) in α-1 antitrypsin mRNA  AAT-1023 21 nt Target: 5′-AAGCGUUUAGGCAUGUUUAAC-3′ (SEQ ID NO: 1907) AAT-1024 21 nt Target: 5′-AGCGUUUAGGCAUGUUUAACA-3′ (SEQ ID NO: 1908) AAT-1404 21 nt Target: 5′-GGGGCCAUGUUUUUAGAGGCC-3′ (SEQ ID NO: 1909) AAT-1461 21 nt Target: 5′-AAACCCUUUGUCUUCUUAAUG-3′ (SEQ ID NO: 1910) AAT-1462 21 nt Target: 5′-AACCCUUUGUCUUCUUAAUGA-3′ (SEQ ID NO: 1911) AAT-1463 21 nt Target: 5′-ACCCUUUGUCUUCUUAAUGAU-3′ (SEQ ID NO: 1912) AAT-1464 21 nt Target: 5′-CCCUUUGUCUUCUUAAUGAUU-3′ (SEQ ID NO: 1913) AAT-1465 21 nt Target: 5′-CCUUUGUCUUCUUAAUGAUUG-3′ (SEQ ID NO: 1914) AAT-1466 21 nt Target: 5′-CUUUGUCUUCUUAAUGAUUGA-3′ (SEQ ID NO: 1915) AAT-1467 21 nt Target: 5′-UUUGUCUUCUUAAUGAUUGAA-3′ (SEQ ID NO: 1916) AAT-1468 21 nt Target: 5′-UUGUCUUCUUAAUGAUUGAAC-3′ (SEQ ID NO: 1917) AAT-1469 21 nt Target: 5′-UGUCUUCUUAAUGAUUGAACA-3′ (SEQ ID NO: 1918) AAT-1470 21 nt Target: 5′-GUCUUCUUAAUGAUUGAACAA-3′ (SEQ ID NO: 1919) AAT-1471 21 nt Target: 5′-UCUUCUUAAUGAUUGAACAAA-3′ (SEQ ID NO: 1920) AAT-1472 21 nt Target: 5′-CUUCUUAAUGAUUGAACAAAA-3′ (SEQ ID NO: 1921) AAT-1473 21 nt Target: 5′-UUCUUAAUGAUUGAACAAAAU-3′ (SEQ ID NO: 1922) AAT-1474 21 nt Target: 5′-UCUUAAUGAUUGAACAAAAUA-3′ (SEQ ID NO: 1923) AAT-1475 21 nt Target: 5′-CUUAAUGAUUGAACAAAAUAC-3′ (SEQ ID NO: 1924) AAT-1476 21 nt Target: 5′-UUAAUGAUUGAACAAAAUACC-3′ (SEQ ID NO: 1925) AAT-1477 21 nt Target: 5′-UAAUGAUUGAACAAAAUACCA-3′ (SEQ ID NO: 1926) AAT-1478 21 nt Target: 5′-AAUGAUUGAACAAAAUACCAA-3′ (SEQ ID NO: 1927) AAT-1479 21 nt Target: 5′-AUGAUUGAACAAAAUACCAAG-3′ (SEQ ID NO: 1928) AAT-1480 21 nt Target: 5′-UGAUUGAACAAAAUACCAAGU-3′ (SEQ ID NO: 1929) AAT-1481 21 nt Target: 5′-GAUUGAACAAAAUACCAAGUC-3′ (SEQ ID NO: 1930) AAT-1482 21 nt Target: 5′-AUUGAACAAAAUACCAAGUCU-3′ (SEQ ID NO: 1931) AAT-1483 21 nt Target: 5′-UUGAACAAAAUACCAAGUCUC-3′ (SEQ ID NO: 1932) AAT-1484 21 nt Target: 5′-UGAACAAAAUACCAAGUCUCC-3′ (SEQ ID NO: 1933) AAT-1485 21 nt Target: 5′-GAACAAAAUACCAAGUCUCCC-3′ (SEQ ID NO: 1934) AAT-1486 21 nt Target: 5′-AACAAAAUACCAAGUCUCCCC-3′ (SEQ ID NO: 1935) AAT-1487 21 nt Target: 5′-ACAAAAUACCAAGUCUCCCCU-3′ (SEQ ID NO: 1936) AAT-1488 21 nt Target: 5′-CAAAAUACCAAGUCUCCCCUC-3′ (SEQ ID NO: 1937) 

TABLE 10  Additional Human Anti-α-1 antitrypsin “Blunt/Blunt” DsiRNTAs  5′-AAGCGUUUAGGCAUGUUUAACAUCCAG-3′ (SEQ ID NO: 1938) 3′-UUCGCAAAUCCGUACAAAUUGUAGGUC-5′ (SEQ ID NO: 1814) AAT-1023 Target: 5′-AAGCGTTTAGGCATGTTTAACATCCAG-3′ (SEQ ID NO: 1845) 5′-AGCGUUUAGGCAUGUUUAACAUCCAGC-3′ (SEQ ID NO: 1939) 3′-UCGCAAAUCCGUACAAAUUGUAGGUCG-5′ (SEQ ID NO: 1815) AAT-1024 Target: 5′-AGCGTTTAGGCATGTTTAACATCCAGC-3′ (SEQ ID NO: 1846) 5′-GGGGCCAUGUUUUUAGAGGCCAUACCC-3′ (SEQ ID NO: 1940) 3′-CCCCGGUACAAAAAUCUCCGGUAUGGG-5′ (SEQ ID NO: 1816) AAT-1404 Target: 5′-GGGGCCATGTTTTTAGAGGCCATACCC-3′ (SEQ ID NO: 1847) 5′-AAACCCUUUGUCUUCUUAAUGAUUGAA-3′ (SEQ ID NO: 1941) 3′-UUUGGGAAACAGAAGAAUUACUAACUU-5′ (SEQ ID NO: 1817) AAT-1461 Target: 5′-AAACCCTTTGTCTTCTTAATGATTGAA-3′ (SEQ ID NO: 1848) 5′-AACCCUUUGUCUUCUUAAUGAUUGAAC-3′ (SEQ ID NO: 1942) 3′-UUGGGAAACAGAAGAAUUACUAACUUG-5′ (SEQ ID NO: 1818) AAT-1462 Target: 5′-AACCCTTTGTCTTCTTAATGATTGAAC-3′ (SEQ ID NO: 1849) 5′-ACCCUUUGUCUUCUUAAUGAUUGAACA-3′ (SEQ ID NO: 1943) 3′-UGGGAAACAGAAGAAUUACUAACUUGU-5′ (SEQ ID NO: 1819) AAT-1463 Target: 5′-ACCCTTTGTCTTCTTAATGATTGAACA-3′ (SEQ ID NO: 1850) 5′-CCCUUUGUCUUCUUAAUGAUUGAACAA-3′ (SEQ ID NO: 1944) 3′-GGGAAACAGAAGAAUUACUAACUUGUU-5′(SEQ ID NO: 1820) AAT-1464 Target: 5′-CCCTTTGTCTTCTTAATGATTGAACAA-3′ (SEQ ID NO: 1851) 5′-CCUUUGUCUUCUUAAUGAUUGAACAAA-3′(SEQ ID NO: 1945) 3′-GGAAACAGAAGAAUUACUAACUUGUUU-5′(SEQ ID NO: 1821) AAT-1465 Target: 5′-CCTTTGTCTTCTTAATGATTGAACAAA-3′ (SEQ ID NO: 1852) 5′-CUUUGUCUUCUUAAUGAUUGAACAAAA-3′ (SEQ ID NO: 1946) 3′-GAAACAGAAGAAUUACUAACUUGUUUU-5′ (SEQ ID NO: 1822) AAT-1466 Target: 5′-CTTTGTCTTCTTAATGATTGAACAAAA-3′ (SEQ ID NO: 1853) 5′-UUUGUCUUCUUAAUGAUUGAACAAAAU-3′ (SEQ ID NO: 1947) 3′-AAACAGAAGAAUUACUAACUUGUUUUA-5′ (SEQ ID NO: 1823) AAT-1467 Target: 5′-TTTGTCTTCTTAATGATTGAACAAAAT-3′ (SEQ ID NO: 1854) 5′-UUGUCUUCUUAAUGAUUGAACAAAAUA-3′ (SEQ ID NO: 1948) 3′-AACAGAAGAAUUACUAACUUGUUUUAU-5′ (SEQ ID NO: 1824) AAT-1468 Target: 5′-TTGTCTTCTTAATGATTGAACAAAATA-3′ (SEQ ID NO: 1855) 5′-UGUCUUCUUAAUGAUUGAACAAAAUAC-3′ (SEQ ID NO: 1949) 3′-ACAGAAGAAUUACUAACUUGUUUUAUG-5′ (SEQ ID NO: 1825) AAT-1469 Target: 5′-TGTCTTCTTAATGATTGAACAAAATAC-3′ (SEQ ID NO: 1856) 5′-GUCUUCUUAAUGAUUGAACAAAAUACC-3′ (SEQ ID NO: 1950) 3′-CAGAAGAAUUACUAACUUGUUUUAUGG-5′ (SEQ ID NO: 1826) AAT-1470 Target: 5′-GTCTTCTTAATGATTGAACAAAATACC-3′ (SEQ ID NO: 1857) 5′-UCUUCUUAAUGAUUGAACAAAAUACCA-3′ (SEQ ID NO: 1951) 3′-AGAAGAAUUACUAACUUGUUUUAUGGU-5′ (SEQ ID NO: 1827) AAT-1471 Target: 5′-TCTTCTTAATGATTGAACAAAATACCA-3′ (SEQ ID NO: 1858) 5′-CUUCUUAAUGAUUGAACAAAAUACCAA-3′(SEQ ID NO: 1952) 3′-GAAGAAUUACUAACUUGUUUUAUGGUU-5′ (SEQ ID NO: 1828) AAT-1472 Target: 5′-CTTCTTAATGATTGAACAAAATACCAA-3′ (SEQ ID NO: 1859) 5′-UUCUUAAUGAUUGAACAAAAUACCAAG-3′ (SEQ ID NO: 1953) 3′-AAGAAUUACUAACUUGUUUUAUGGUUC-5′ (SEQ ID NO: 1829) AAT-1473 Target: 5′-TTCTTAATGATTGAACAAAATACCAAG-3′ (SEQ ID NO: 1860) 5′-UCUUAAUGAUUGAACAAAAUACCAAGU-3′ (SEQ ID NO: 1954) 3′-AGAAUUACUAACUUGUUUUAUGGUUCA-5′ (SEQ ID NO: 1830) AAT-1474 Target: 5′-TCTTAATGATTGAACAAAATACCAAGT-3′ (SEQ ID NO: 1861) 5′-CUUAAUGAUUGAACAAAAUACCAAGUC-3′ (SEQ ID NO: 1955) 3′-GAAUUACUAACUUGUUUUAUGGUUCAG-5′ (SEQ ID NO: 1831) AAT-1475 Target: 5′-CTTAATGATTGAACAAAATACCAAGTC-3′ (SEQ ID NO: 1862) 5′-UUAAUGAUUGAACAAAAUACCAAGUCU-3′ (SEQ ID NO: 1956) 3′-AAUUACUAACUUGUUUUAUGGUUCAGA-5′ (SEQ ID NO: 1832) AAT-1476 Target: 5′-TTAATGATTGAACAAAATACCAAGTCT-3′ (SEQ ID NO: 1863) 5′-UAAUGAUUGAACAAAAUACCAAGUCUC-3′ (SEQ ID NO: 1957) 3′-AUUACUAACUUGUUUUAUGGUUCAGAG-5′ (SEQ ID NO: 1833) AAT-1477 Target: 5′-TAATGATTGAACAAAATACCAAGTCTC-3′ (SEQ ID NO: 1864) 5′-AAUGAUUGAACAAAAUACCAAGUCUCC-3′ (SEQ ID NO: 1958) 3′-UUACUAACUUGUUUUAUGGUUCAGAGG-5′ (SEQ ID NO: 1834) AAT-1478 Target: 5′-AATGATTGAACAAAATACCAAGTCTCC-3′ (SEQ ID NO: 1865) 5′-AUGAUUGAACAAAAUACCAAGUCUCCC-3′ (SEQ ID NO: 1959) 3′-UACUAACUUGUUUUAUGGUUCAGAGGG-5′ (SEQ ID NO: 1835) AAT-1479 Target: 5′-ATGATTGAACAAAATACCAAGTCTCCC-3′ (SEQ ID NO: 1866) 5′-UGAUUGAACAAAAUACCAAGUCUCCCC-3′ (SEQ ID NO: 1960) 3′-ACUAACUUGUUUUAUGGUUCAGAGGGG-5′ (SEQ ID NO: 1836) AAT-1480 Target: 5′-TGATTGAACAAAATACCAAGTCTCCCC-3′ (SEQ ID NO: 1867) 5′-GAUUGAACAAAAUACCAAGUCUCCCCU-3′ (SEQ ID NO: 1961) 3′-CUAACUUGUUUUAUGGUUCAGAGGGGA-5′ (SEQ ID NO: 1837) AAT-1481 Target: 5′-GATTGAACAAAATACCAAGTCTCCCCT-3′ (SEQ ID NO: 1868) 5′-AUUGAACAAAAUACCAAGUCUCCCCUC-3′ (SEQ ID NO: 1962) 3′-UAACUUGUUUUAUGGUUCAGAGGGGAG-5′ (SEQ ID NO: 1838) AAT-1482 Target: 5′-ATTGAACAAAATACCAAGTCTCCCCTC-3′ (SEQ ID NO: 1869) 5′-UUGAACAAAAUACCAAGUCUCCCCUCU-3′ (SEQ ID NO: 1963) 3′-AACUUGUUUUAUGGUUCAGAGGGGAGA-5′ (SEQ ID NO: 1839) AAT-1483 Target: 5′-TTGAACAAAATACCAAGTCTCCCCTCT-3′ (SEQ ID NO: 1870) 5′-UGAACAAAAUACCAAGUCUCCCCUCUU-3′ (SEQ ID NO: 1964) 3′-ACUUGUUUUAUGGUUCAGAGGGGAGAA-5′ (SEQ ID NO: 1840) AAT-1484 Target: 5′-TGAACAAAATACCAAGTCTCCCCTCTT-3′ (SEQ ID NO: 1871) 5′-GAACAAAAUACCAAGUCUCCCCUCUUC-3′ (SEQ ID NO: 1965) 3′-CUUGUUUUAUGGUUCAGAGGGGAGAAG-5′ (SEQ ID NO: 1841) AAT-1485 Target: 5′-GAACAAAATACCAAGTCTCCCCTCTTC-3′ (SEQ ID NO: 1872) 5′-AACAAAAUACCAAGUCUCCCCUCUUCA-3′ (SEQ ID NO: 1966) 3′-UUGUUUUAUGGUUCAGAGGGGAGAAGU-5′ (SEQ ID NO: 1842) AAT-1486 Target: 5′-AACAAAATACCAAGTCTCCCCTCTTCA-3′ (SEQ ID NO: 1873) 5′-ACAAAAUACCAAGUCUCCCCUCUUCAU-3′ (SEQ ID NO: 1967) 3′-UGUUUUAUGGUUCAGAGGGGAGAAGUA-5′ (SEQ ID NO: 1843) AAT-1487 Target: 5′-ACAAAATACCAAGTCTCCCCTCTTCAT-3′ (SEQ ID NO: 1874) 5′-CAAAAUACCAAGUCUCCCCUCUUCAUG-3′ (SEQ ID NO: 1968) 3′-GUUUUAUGGUUCAGAGGGGAGAAGUAC-5′ (SEQ ID NO: 1844) AAT-1488 Target: 5′-CAAAATACCAAGTCTCCCCTCTTCATG-3′ (SEQ ID NO: 1875) 

TABLE 11  Additional DsiRNA Component 19 Nucleotide Target Sequencesin α-1 antitrypsin mRNA  AAT-1023 19 nt Target #1: 5′-GCGUUUAGGCAUGUUUAAC-3′ (SEQ ID NO: 1969) AAT-1023 19 nt Target #2: 5′-AGCGUUUAGGCAUGUUUAA-3′ (SEQ ID NO: 2000) AAT-1023 19 nt Target #3: 5′-AAGCGUUUAGGCAUGUUUA-3′ (SEQ ID NO: 2031) AAT-1024 19 nt Target #1: 5′-CGUUUAGGCAUGUUUAACA-3′ (SEQ ID NO: 1970) AAT-1024 19 nt Target #2: 5′-GCGUUUAGGCAUGUUUAAC-3′ (SEQ ID NO: 2001) AAT-1024 19 nt Target #3: 5′-AGCGUUUAGGCAUGUUUAA-3′ (SEQ ID NO: 2032) AAT-1404 19 nt Target #1: 5′-GGCCAUGUUUUUAGAGGCC-3′ (SEQ ID NO: 1971) AAT-1404 19 nt Target #2: 5′-GGGCCAUGUUUUUAGAGGC-3′ (SEQ ID NO: 2002) AAT-1404 19 nt Target #3: 5′-GGGGCCAUGUUUUUAGAGG-3′ (SEQ ID NO: 2033) AAT-1461 19 nt Target #1: 5′-ACCCUUUGUCUUCUUAAUG-3′ (SEQ ID NO: 1972) AAT-1461 19 nt Target #2: 5′-AACCCUUUGUCUUCUUAAU-3′ (SEQ ID NO: 2003) AAT-1461 19 nt Target #3: 5′-AAACCCUUUGUCUUCUUAA-3′ (SEQ ID NO: 2034) AAT-1462 19 nt Target #1: 5′-CCCUUUGUCUUCUUAAUGA-3′ (SEQ ID NO: 1973) AAT-1462 19 nt Target #2: 5′-ACCCUUUGUCUUCUUAAUG-3′ (SEQ ID NO: 2004) AAT-1462 19 nt Target #3: 5′-AACCCUUUGUCUUCUUAAU-3′ (SEQ ID NO: 2035) AAT-1463 19 nt Target #1: 5′-CCUUUGUCUUCUUAAUGAU-3′ (SEQ ID NO: 1974) AAT-1463 19 nt Target #2: 5′-CCCUUUGUCUUCUUAAUGA-3′ (SEQ ID NO: 2005) AAT-1463 19 nt Target #3: 5′-ACCCUUUGUCUUCUUAAUG-3′ (SEQ ID NO: 2036) AAT-1464 19 nt Target #1: 5′-CUUUGUCUUCUUAAUGAUU-3′ (SEQ ID NO: 1975) AAT-1464 19 nt Target #2: 5′-CCUUUGUCUUCUUAAUGAU-3′ (SEQ ID NO: 2006) AAT-1464 19 nt Target #3: 5′-CCCUUUGUCUUCUUAAUGA-3′ (SEQ ID NO: 2037) AAT-1465 19 nt Target #1: 5′-UUUGUCUUCUUAAUGAUUG-3′ (SEQ ID NO: 1976) AAT-1465 19 nt Target #2: 5′-CUUUGUCUUCUUAAUGAUU-3′ (SEQ ID NO: 2007) AAT-1465 19 nt Target #3: 5′-CCUUUGUCUUCUUAAUGAU-3′ (SEQ ID NO: 2038) AAT-1466 19 nt Target #1: 5′-UUGUCUUCUUAAUGAUUGA-3′ (SEQ ID NO: 1977) AAT-1466 19 nt Target #2: 5′-UUUGUCUUCUUAAUGAUUG-3′ (SEQ ID NO: 2008) AAT-1466 19 nt Target #3: 5′-CUUUGUCUUCUUAAUGAUU-3′ (SEQ ID NO: 2039) AAT-1467 19 nt Target #1: 5′-UGUCUUCUUAAUGAUUGAA-3′ (SEQ ID NO: 1978) AAT-1467 19 nt Target #2: 5′-UUGUCUUCUUAAUGAUUGA-3′ (SEQ ID NO: 2009) AAT-1467 19 nt Target #3: 5′-UUUGUCUUCUUAAUGAUUG-3′ (SEQ ID NO: 2040) AAT-1468 19 nt Target #1: 5′-GUCUUCUUAAUGAUUGAAC-3′ (SEQ ID NO: 1979) AAT-1468 19 nt Target #2: 5′-UGUCUUCUUAAUGAUUGAA-3′ (SEQ ID NO: 2010) AAT-1468 19 nt Target #3: 5′-UUGUCUUCUUAAUGAUUGA-3′ (SEQ ID NO: 2041) AAT-1469 19 nt Target #1: 5′-UCUUCUUAAUGAUUGAACA-3′ (SEQ ID NO: 1980) AAT-1469 19 nt Target #2: 5′-GUCUUCUUAAUGAUUGAAC-3′ (SEQ ID NO: 2011) AAT-1469 19 nt Target #3: 5′-UGUCUUCUUAAUGAUUGAA-3′ (SEQ ID NO: 2042) AAT-1470 19 nt Target #1: 5′-CUUCUUAAUGAUUGAACAA-3′ (SEQ ID NO: 1981) AAT-1470 19 nt Target #2: 5′-UCUUCUUAAUGAUUGAACA-3′ (SEQ ID NO: 2012) AAT-1470 19 nt Target #3: 5′-GUCUUCUUAAUGAUUGAAC-3′ (SEQ ID NO: 2043) AAT-1471 19 nt Target #1: 5′-UUCUUAAUGAUUGAACAAA-3′ (SEQ ID NO: 1982) AAT-1471 19 nt Target #2: 5′-CUUCUUAAUGAUUGAACAA-3′ (SEQ ID NO: 2013) AAT-1471 19 nt Target #3: 5′-UCUUCUUAAUGAUUGAACA-3′ (SEQ ID NO: 2044) AAT-1472 19 nt Target #1: 5′-UCUUAAUGAUUGAACAAAA-3′ (SEQ ID NO: 1983) AAT-1472 19 nt Target #2: 5′-UUCUUAAUGAUUGAACAAA-3′ (SEQ ID NO: 2014) AAT-1472 19 nt Target #3: 5′-CUUCUUAAUGAUUGAACAA-3′ (SEQ ID NO: 2045) AAT-1473 19 nt Target #1: 5′-CUUAAUGAUUGAACAAAAU-3′ (SEQ ID NO: 1984) AAT-1473 19 nt Target #2: 5′-UCUUAAUGAUUGAACAAAA-3′ (SEQ ID NO: 2015) AAT-1473 19 nt Target #3: 5′-UUCUUAAUGAUUGAACAAA-3′ (SEQ ID NO: 2046) AAT-1474 19 nt Target #1: 5′-UUAAUGAUUGAACAAAAUA-3′ (SEQ ID NO: 1985) AAT-1474 19 nt Target #2: 5′-CUUAAUGAUUGAACAAAAU-3′ (SEQ ID NO: 2016) AAT-1474 19 nt Target #3: 5′-UCUUAAUGAUUGAACAAAA-3′ (SEQ ID NO: 2047) AAT-1475 19 nt Target #1: 5′-UAAUGAUUGAACAAAAUAC-3′ (SEQ ID NO: 1986) AAT-1475 19 nt Target #2: 5′-UUAAUGAUUGAACAAAAUA-3′ (SEQ ID NO: 2017) AAT-1475 19 nt Target #3: 5′-CUUAAUGAUUGAACAAAAU-3′ (SEQ ID NO: 2048) AAT-1476 19 nt Target #1: 5′-AAUGAUUGAACAAAAUACC-3′ (SEQ ID NO: 1987) AAT-1476 19 nt Target #2: 5′-UAAUGAUUGAACAAAAUAC-3′ (SEQ ID NO: 2018) AAT-1476 19 nt Target #3: 5′-UUAAUGAUUGAACAAAAUA-3′ (SEQ ID NO: 2049) AAT-1477 19 nt Target #1: 5′-AUGAUUGAACAAAAUACCA-3′ (SEQ ID NO: 1988) AAT-1477 19 nt Target #2: 5′-AAUGAUUGAACAAAAUACC-3′ (SEQ ID NO: 2019) AAT-1477 19 nt Target #3: 5′-UAAUGAUUGAACAAAAUAC-3′ (SEQ ID NO: 2050) AAT-1478 19 nt Target #1: 5′-UGAUUGAACAAAAUACCAA-3′ (SEQ ID NO: 1989) AAT-1478 19 nt Target #2: 5′-AUGAUUGAACAAAAUACCA-3′ (SEQ ID NO: 2020) AAT-1478 19 nt Target #3: 5′-AAUGAUUGAACAAAAUACC-3′ (SEQ ID NO: 2051) AAT-1479 19 nt Target #1: 5′-GAUUGAACAAAAUACCAAG-3′ (SEQ ID NO: 1990) AAT-1479 19 nt Target #2: 5′-UGAUUGAACAAAAUACCAA-3′ (SEQ ID NO: 2021) AAT-1479 19 nt Target #3: 5′-AUGAUUGAACAAAAUACCA-3′ (SEQ ID NO: 2052) AAT-1480 19 nt Target #1: 5′-AUUGAACAAAAUACCAAGU-3′ (SEQ ID NO: 1991) AAT-1480 19 nt Target #2: 5′-GAUUGAACAAAAUACCAAG-3′ (SEQ ID NO: 2022) AAT-1480 19 nt Target #3: 5′-UGAUUGAACAAAAUACCAA-3′ (SEQ ID NO: 2053) AAT-1481 19 nt Target #1: 5′-UUGAACAAAAUACCAAGUC-3′ (SEQ ID NO: 1992) AAT-1481 19 nt Target #2: 5′-AUUGAACAAAAUACCAAGU-3′ (SEQ ID NO: 2023) AAT-1481 19 nt Target #3: 5′-GAUUGAACAAAAUACCAAG-3′ (SEQ ID NO: 2054) AAT-1482 19 nt Target #1: 5′-UGAACAAAAUACCAAGUCU-3′ (SEQ ID NO: 1993) AAT-1482 19 nt Target #2: 5′-UUGAACAAAAUACCAAGUC-3′ (SEQ ID NO: 2024) AAT-1482 19 nt Target #3: 5′-AUUGAACAAAAUACCAAGU-3′ (SEQ ID NO: 2055) AAT-1483 19 nt Target #1: 5′-GAACAAAAUACCAAGUCUC-3′ (SEQ ID NO: 1994) AAT-1483 19 nt Target #2: 5′-UGAACAAAAUACCAAGUCU-3′ (SEQ ID NO: 2025) AAT-1483 19 nt Target #3: 5′-UUGAACAAAAUACCAAGUC-3′ (SEQ ID NO: 2056) AAT-1484 19 nt Target #1: 5′-AACAAAAUACCAAGUCUCC-3′ (SEQ ID NO: 1995) AAT-1484 19 nt Target #2: 5′-GAACAAAAUACCAAGUCUC-3′ (SEQ ID NO: 2026) AAT-1484 19 nt Target #3: 5′-UGAACAAAAUACCAAGUCU-3′ (SEQ ID NO: 2057) AAT-1485 19 nt Target #1: 5′-ACAAAAUACCAAGUCUCCC-3′ (SEQ ID NO: 1996) AAT-1485 19 nt Target #2: 5′-AACAAAAUACCAAGUCUCC-3′ (SEQ ID NO: 2027) AAT-1485 19 nt Target #3: 5′-GAACAAAAUACCAAGUCUC-3′ (SEQ ID NO: 2058) AAT-1486 19 nt Target #1: 5′-CAAAAUACCAAGUCUCCCC-3′ (SEQ ID NO: 1997) AAT-1486 19 nt Target #2: 5′-ACAAAAUACCAAGUCUCCC-3′ (SEQ ID NO: 2028) AAT-1486 19 nt Target #3: 5′-AACAAAAUACCAAGUCUCC-3′ (SEQ ID NO: 2059) AAT-1487 19 nt Target #1: 5′-AAAAUACCAAGUCUCCCCU-3′ (SEQ ID NO: 1998) AAT-1487 19 nt Target #2: 5′-CAAAAUACCAAGUCUCCCC-3′ (SEQ ID NO: 2029) AAT-1487 19 nt Target #3: 5′-ACAAAAUACCAAGUCUCCC-3′ (SEQ ID NO: 2060) AAT-1488 19 nt Target #1: 5′-AAAUACCAAGUCUCCCCUC-3′ (SEQ ID NO: 1999) AAT-1488 19 nt Target #2: 5′-AAAAUACCAAGUCUCCCCU-3′ (SEQ ID NO: 2030) AAT-1488 19 nt Target #3: 5′-CAAAAUACCAAGUCUCCCC-3′ (SEQ ID NO: 2061)

TABLE 12  Further Human Anti-α-1 antitrypsin DsiRNA Agents (Asymmetrics)  5′-CCAGGGAGAUGCUGCCCAGAAGAca-3′ (SEQ ID NO: 2062) 3′-GGGGUCCCUCUACGACGGGUCUUCUGU-5′ (SEQ ID NO: 2202) AAT-366 Target: 5′-CCCCAGGGAGATGCTGCCCAGAAGACA-3′ (SEQ ID NO: 2342) 5′-CAGGGAGAUGCUGCCCAGAAGACag-3′ (SEQ ID NO: 2063) 3′-GGGUCCCUCUACGACGGGUCUUCUGUC-5′ (SEQ ID NO: 2203) AAT-367 Target: 5′-CCCAGGGAGATGCTGCCCAGAAGACAG-3′ (SEQ ID NO: 2343) 5′-AGGGAGAUGCUGCCCAGAAGACAga-3′ (SEQ ID NO: 2064) 3′-GGUCCCUCUACGACGGGUCUUCUGUCU-5′ (SEQ ID NO: 2204) AAT-368 Target: 5′-CCAGGGAGATGCTGCCCAGAAGACAGA-3′ (SEQ ID NO: 2344) 5′-GGGAGAUGCUGCCCAGAAGACAGat-3′ (SEQ ID NO: 2065) 3′-GUCCCUCUACGACGGGUCUUCUGUCUA-5′ (SEQ ID NO: 2205) AAT-369 Target: 5′-CAGGGAGATGCTGCCCAGAAGACAGAT-3′ (SEQ ID NO: 2345) 5′-GGAGAUGCUGCCCAGAAGACAGAta-3′ (SEQ ID NO: 2066) 3′-UCCCUCUACGACGGGUCUUCUGUCUAU-5′ (SEQ ID NO: 2206) AAT-370 Target: 5′-AGGGAGATGCTGCCCAGAAGACAGATA-3′ (SEQ ID NO: 2346) 5′-GAGAUGCUGCCCAGAAGACAGAUac-3′ (SEQ ID NO: 2067) 3′-CCCUCUACGACGGGUCUUCUGUCUAUG-5′ (SEQ ID NO: 2207) AAT-371 Target: 5′-GGGAGATGCTGCCCAGAAGACAGATAC-3′ (SEQ ID NO: 2347) 5′-GAUACAUCCCACCAUGAUCAGGAtc-3′ (SEQ ID NO: 2068) 3′-GUCUAUGUAGGGUGGUACUAGUCCUAG-5′ (SEQ ID NO: 2208) AAT-391 Target: 5′-CAGATACATCCCACCATGATCAGGATC-3′ (SEQ ID NO: 2348) 5′-AUACAUCCCACCAUGAUCAGGAUca-3′ (SEQ ID NO: 2069) 3′-UCUAUGUAGGGUGGUACUAGUCCUAGU-5′ (SEQ ID NO: 2209) AAT-392 Target: 5′-AGATACATCCCACCATGATCAGGATCA-3′ (SEQ ID NO: 2349) 5′-UACAUCCCACCAUGAUCAGGAUCac-3′ (SEQ ID NO: 2070) 3′-CUAUGUAGGGUGGUACUAGUCCUAGUG-5′ (SEQ ID NO: 2210) AAT-393 Target: 5′-GATACATCCCACCATGATCAGGATCAC-3′ (SEQ ID NO: 2350) 5′-ACAUCCCACCAUGAUCAGGAUCAcc-3′ (SEQ ID NO: 2071) 3′-UAUGUAGGGUGGUACUAGUCCUAGUGG-5′ (SEQ ID NO: 2211) AAT-394 Target: 5′-ATACATCCCACCATGATCAGGATCACC-3′ (SEQ ID NO: 2351) 5′-ACCAGUCCAACAGCACCAAUAUCtt-3′ (SEQ ID NO: 2072) 3′-UGUGGUCAGGUUGUCGUGGUUAUAGAA-5′ (SEQ ID NO: 2212) AAT-485 Target: 5′-ACACCAGTCCAACAGCACCAATATCTT-3′ (SEQ ID NO: 2352) 5′-CCAGUCCAACAGCACCAAUAUCUtc-3′ (SEQ ID NO: 2073) 3′-GUGGUCAGGUUGUCGUGGUUAUAGAAG-5′ (SEQ ID NO: 2213) AAT-486 Target: 5′-CACCAGTCCAACAGCACCAATATCTTC-3′ (SEQ ID NO: 2353) 5′-CAGUCCAACAGCACCAAUAUCUUct-3′ (SEQ ID NO: 2074) 3′-UGGUCAGGUUGUCGUGGUUAUAGAAGA-5′ (SEQ ID NO: 2214) AAT-487 Target: 5′-ACCAGTCCAACAGCACCAATATCTTCT-3′ (SEQ ID NO: 2354) 5′-AGUCCAACAGCACCAAUAUCUUCtt-3′ (SEQ ID NO: 2075) 3′-GGUCAGGUUGUCGUGGUUAUAGAAGAA-5′ (SEQ ID NO: 2215) AAT-488 Target: 5′-CCAGTCCAACAGCACCAATATCTTCTT-3′ (SEQ ID NO: 2355) 5′-GUCCAACAGCACCAAUAUCUUCUtc-3′ (SEQ ID NO: 2076) 3′-GUCAGGUUGUCGUGGUUAUAGAAGAAG-5′(SEQ ID NO: 2216) AAT-489 Target: 5′-CAGTCCAACAGCACCAATATCTTCTTC-3′ (SEQ ID NO: 2356) 5′-UCCAACAGCACCAAUAUCUUCUUct-3′ (SEQ ID NO: 2077) 3′-UCAGGUUGUCGUGGUUAUAGAAGAAGA-5′ (SEQ ID NO: 2217) AAT-490 Target: 5′-AGTCCAACAGCACCAATATCTTCTTCT-3′ (SEQ ID NO: 2357) 5′-CCAACAGCACCAAUAUCUUCUUCtc-3′ (SEQ ID NO: 2078) 3′-CAGGUUGUCGUGGUUAUAGAAGAAGAG-5′ (SEQ ID NO: 2218) AAT-491 Target: 5′-GTCCAACAGCACCAATATCTTCTTCTC-3′ (SEQ ID NO: 2358) 5′-CAACAGCACCAAUAUCUUCUUCUcc-3′ (SEQ ID NO: 2079) 3′-AGGUUGUCGUGGUUAUAGAAGAAGAGG-5′ (SEQ ID NO: 2219) AAT-492 Target: 5′-TCCAACAGCACCAATATCTTCTTCTCC-3′ (SEQ ID NO: 2359) 5′-AACAGCACCAAUAUCUUCUUCUCcc-3′ (SEQ ID NO: 2080) 3′-GGUUGUCGUGGUUAUAGAAGAAGAGGG-5′ (SEQ ID NO: 2220) AAT-493 Target: 5′-CCAACAGCACCAATATCTTCTTCTCCC-3′ (SEQ ID NO: 2360) 5′-ACAGCACCAAUAUCUUCUUCUCCcc-3′ (SEQ ID NO: 2081) 3′-GUUGUCGUGGUUAUAGAAGAAGAGGGG-5′ (SEQ ID NO: 2221) AAT-494 Target: 5′-CAACAGCACCAATATCTTCTTCTCCCC-3′ (SEQ ID NO: 2361) 5′-CAGCACCAAUAUCUUCUUCUCCCca-3′ (SEQ ID NO: 2082) 3′-UUGUCGUGGUUAUAGAAGAAGAGGGGU-5′ (SEQ ID NO: 2222) AAT-495 Target: 5′-AACAGCACCAATATCTTCTTCTCCCCA-3′ (SEQ ID NO: 2362) 5′-AGCACCAAUAUCUUCUUCUCCCCag-3′ (SEQ ID NO: 2083) 3′-UGUCGUGGUUAUAGAAGAAGAGGGGUC-5′ (SEQ ID NO: 2223) AAT-496 Target: 5′-ACAGCACCAATATCTTCTTCTCCCCAG-3′ (SEQ ID NO: 2363) 5′-GCACCAAUAUCUUCUUCUCCCCAgt-3′ (SEQ ID NO: 2084) 3′-GUCGUGGUUAUAGAAGAAGAGGGGUCA-5′ (SEQ ID NO: 2224) AAT-497 Target: 5′-CAGCACCAATATCTTCTTCTCCCCAGT-3′ (SEQ ID NO: 2364) 5′-CACCAAUAUCUUCUUCUCCCCAGtg-3′ (SEQ ID NO: 2085) 3′-UCGUGGUUAUAGAAGAAGAGGGGUCAC-5′ (SEQ ID NO: 2225) AAT-498 Target: 5′-AGCACCAATATCTTCTTCTCCCCAGTG-3′ (SEQ ID NO: 2365) 5′-ACCAAUAUCUUCUUCUCCCCAGUga-3′ (SEQ ID NO: 2086) 3′-CGUGGUUAUAGAAGAAGAGGGGUCACU-5′ (SEQ ID NO: 2226) AAT-499 Target: 5′-GCACCAATATCTTCTTCTCCCCAGTGA-3′ (SEQ ID NO: 2366) 5′-CCCAGUGAGCAUCGCUACAGCCUtt-3′ (SEQ ID NO: 2087) 3′-AGGGGUCACUCGUAGCGAUGUCGGAAA-5′ (SEQ ID NO: 2227) AAT-516 Target: 5′-TCCCCAGTGAGCATCGCTACAGCCTTT-3′ (SEQ ID NO: 2367) 5′-CCAGUGAGCAUCGCUACAGCCUUtg-3′ (SEQ ID NO: 2088) 3′-GGGGUCACUCGUAGCGAUGUCGGAAAC-5′ (SEQ ID NO: 2228) AAT-517 Target: 5′-CCCCAGTGAGCATCGCTACAGCCTTTG-3′ (SEQ ID NO: 2368) 5′-CAGUGAGCAUCGCUACAGCCUUUgc-3′ (SEQ ID NO: 2089) 3′-GGGUCACUCGUAGCGAUGUCGGAAACG-5′ (SEQ ID NO: 2229) AAT-518 Target: 5′-CCCAGTGAGCATCGCTACAGCCTTTGC-3′ (SEQ ID NO: 2369) 5′-AGUGAGCAUCGCUACAGCCUUUGca-3′ (SEQ ID NO: 2090) 3′-GGUCACUCGUAGCGAUGUCGGAAACGU-5′ (SEQ ID NO: 2230) AAT-519 Target: 5′-CCAGTGAGCATCGCTACAGCCTTTGCA-3′ (SEQ ID NO: 2370) 5′-GUGAGCAUCGCUACAGCCUUUGCaa-3′ (SEQ ID NO: 2091) 3′-GUCACUCGUAGCGAUGUCGGAAACGUU-5′ (SEQ ID NO: 2231) AAT-520 Target: 5′-CAGTGAGCATCGCTACAGCCTTTGCAA-3′ (SEQ ID NO: 2371) 5′-UGAGCAUCGCUACAGCCUUUGCAat-3′ (SEQ ID NO: 2092) 3′-UCACUCGUAGCGAUGUCGGAAACGUUA-5′ (SEQ ID NO: 2232) AAT-521 Target: 5′-AGTGAGCATCGCTACAGCCTTTGCAAT-3′ (SEQ ID NO: 2372) 5′-GAGCAUCGCUACAGCCUUUGCAAtg-3′ (SEQ ID NO: 2093) 3′-CACUCGUAGCGAUGUCGGAAACGUUAC-5′ (SEQ ID NO: 2233) AAT-522 Target: 5′-GTGAGCATCGCTACAGCCTTTGCAATG-3′ (SEQ ID NO: 2373) 5′-AGCAUCGCUACAGCCUUUGCAAUgc-3′ (SEQ ID NO: 2094) 3′-ACUCGUAGCGAUGUCGGAAACGUUACG-5′ (SEQ ID NO: 2234) AAT-523 Target: 5′-TGAGCATCGCTACAGCCTTTGCAATGC-3′ (SEQ ID NO: 2374) 5′-GCAUCGCUACAGCCUUUGCAAUGct-3′ (SEQ ID NO: 2095) 3′-CUCGUAGCGAUGUCGGAAACGUUACGA-5′ (SEQ ID NO: 2235) AAT-524 Target: 5′-GAGCATCGCTACAGCCTTTGCAATGCT-3′ (SEQ ID NO: 2375) 5′-CAUCGCUACAGCCUUUGCAAUGCtc-3′ (SEQ ID NO: 2096) 3′-UCGUAGCGAUGUCGGAAACGUUACGAG-5′ (SEQ ID NO: 2236) AAT-525 Target: 5′-AGCATCGCTACAGCCTTTGCAATGCTC-3′ (SEQ ID NO: 2376) 5′-AUCGCUACAGCCUUUGCAAUGCUct-3′ (SEQ ID NO: 2097) 3′-CGUAGCGAUGUCGGAAACGUUACGAGA-5′ (SEQ ID NO: 2237) AAT-526 Target: 5′-GCATCGCTACAGCCTTTGCAATGCTCT-3′ (SEQ ID NO: 2377) 5′-UCGCUACAGCCUUUGCAAUGCUCtc-3′ (SEQ ID NO: 2098) 3′-GUAGCGAUGUCGGAAACGUUACGAGAG-5′ (SEQ ID NO: 2238) AAT-527 Target: 5′-CATCGCTACAGCCTTTGCAATGCTCTC-3′ (SEQ ID NO: 2378) 5′-CGCUACAGCCUUUGCAAUGCUCUcc-3′ (SEQ ID NO: 2099) 3′-UAGCGAUGUCGGAAACGUUACGAGAGG-5′ (SEQ ID NO: 2239) AAT-528 Target: 5′-ATCGCTACAGCCTTTGCAATGCTCTCC-3′ (SEQ ID NO: 2379) 5′-GCUACAGCCUUUGCAAUGCUCUCcc-3′ (SEQ ID NO: 2100) 3′-AGCGAUGUCGGAAACGUUACGAGAGGG-5′ (SEQ ID NO: 2240) AAT-529 Target: 5′-TCGCTACAGCCTTTGCAATGCTCTCCC-3′ (SEQ ID NO: 2380) 5′-CUACAGCCUUUGCAAUGCUCUCCct-3′ (SEQ ID NO: 2101) 3′-GCGAUGUCGGAAACGUUACGAGAGGGA-5′ (SEQ ID NO: 2241) AAT-530 Target: 5′-CGCTACAGCCTTTGCAATGCTCTCCCT-3′ (SEQ ID NO: 2381) 5′-UACAGCCUUUGCAAUGCUCUCCCtg-3′ (SEQ ID NO: 2102) 3′-CGAUGUCGGAAACGUUACGAGAGGGAC-5′ (SEQ ID NO: 2242) AAT-531 Target: 5′-GCTACAGCCTTTGCAATGCTCTCCCTG-3′ (SEQ ID NO: 2382) 5′-CCUGGGGACCAAGGCUGACACUCac-3′ (SEQ ID NO: 2103) 3′-AGGGACCCCUGGUUCCGACUGUGAGUG-5′ (SEQ ID NO: 2243) AAT-552 Target: 5′-TCCCTGGGGACCAAGGCTGACACTCAC-3′ (SEQ ID NO: 2383) 5′-GGGACCAAGGCUGACACUCACGAtg-3′ (SEQ ID NO: 2104) 3′-ACCCCUGGUUCCGACUGUGAGUGCUAC-5′ (SEQ ID NO: 2244) AAT-556 Target: 5′-TGGGGACCAAGGCTGACACTCACGATG-3′ (SEQ ID NO: 2384) 5′-GGACCAAGGCUGACACUCACGAUga-3′ (SEQ ID NO: 2105) 3′-CCCCUGGUUCCGACUGUGAGUGCUACU-5′ (SEQ ID NO: 2245) AAT-557 Target: 5′-GGGGACCAAGGCTGACACTCACGATGA-3′ (SEQ ID NO: 2385) 5′-GACCAAGGCUGACACUCACGAUGaa-3′ (SEQ ID NO: 2106) 3′-CCCUGGUUCCGACUGUGAGUGCUACUU-5′ (SEQ ID NO: 2246) AAT-558 Target: 5′-GGGACCAAGGCTGACACTCACGATGAA-3′ (SEQ ID NO: 2386) 5′-UGAAAUCCUGGAGGGCCUGAAUUtc-3′ (SEQ ID NO: 2107) 3′-CUACUUUAGGACCUCCCGGACUUAAAG-5′ (SEQ ID NO: 2247) AAT-579 Target: 5′-GATGAAATCCTGGAGGGCCTGAATTTC-3′ (SEQ ID NO: 2387) 5′-GAAAUCCUGGAGGGCCUGAAUUUca-3′ (SEQ ID NO: 2108) 3′-UACUUUAGGACCUCCCGGACUUAAAGU-5′ (SEQ ID NO: 2248) AAT-580 Target: 5′-ATGAAATCCTGGAGGGCCTGAATTTCA-3′ (SEQ ID NO: 2388) 5′-UCCAUGAAGGCUUCCAGGAACUCct-3′ (SEQ ID NO: 2109) 3′-CUAGGUACUUCCGAAGGUCCUUGAGGA-5′ (SEQ ID NO: 2249) AAT-632 Target: 5′-GATCCATGAAGGCTTCCAGGAACTCCT-3′ (SEQ ID NO: 2389) 5′-CCAUGAAGGCUUCCAGGAACUCCtc-3′ (SEQ ID NO: 2110) 3′-UAGGUACUUCCGAAGGUCCUUGAGGAG-5′ (SEQ ID NO: 2250) AAT-633 Target: 5′-ATCCATGAAGGCTTCCAGGAACTCCTC-3′ (SEQ ID NO: 2390) 5′-GGACACCGAAGAGGCCAAGAAACag-3′ (SEQ ID NO: 2111) 3′-CCCCUGUGGCUUCUCCGGUUCUUUGUC-5′ (SEQ ID NO: 2251) AAT-801 Target: 5′-GGGGACACCGAAGAGGCCAAGAAACAG-3′ (SEQ ID NO: 2391) 5′-GACACCGAAGAGGCCAAGAAACAga-3′ (SEQ ID NO: 2112) 3′-CCCUGUGGCUUCUCCGGUUCUUUGUCU-5′ (SEQ ID NO: 2252) AAT-802 Target: 5′-GGGACACCGAAGAGGCCAAGAAACAGA-3′ (SEQ ID NO: 2392) 5′-ACACCGAAGAGGCCAAGAAACAGat-3′ (SEQ ID NO: 2113) 3′-CCUGUGGCUUCUCCGGUUCUUUGUCUA-5′ (SEQ ID NO: 2253) AAT-803 Target: 5′-GGACACCGAAGAGGCCAAGAAACAGAT-3′ (SEQ ID NO: 2393) 5′-CACCGAAGAGGCCAAGAAACAGAtc-3′ (SEQ ID NO: 2114) 3′-CUGUGGCUUCUCCGGUUCUUUGUCUAG-5′ (SEQ ID NO: 2254) AAT-804 Target: 5′-GACACCGAAGAGGCCAAGAAACAGATC-3′ (SEQ ID NO: 2394) 5′-ACCGAAGAGGCCAAGAAACAGAUca-3′ (SEQ ID NO: 2115) 3′-UGUGGCUUCUCCGGUUCUUUGUCUAGU-5′ (SEQ ID NO: 2255) AAT-805 Target: 5′-ACACCGAAGAGGCCAAGAAACAGATCA-3′ (SEQ ID NO: 2395) 5′-CCGAAGAGGCCAAGAAACAGAUCaa-3′ (SEQ ID NO: 2116) 3′-GUGGCUUCUCCGGUUCUUUGUCUAGUU-5′ (SEQ ID NO: 2256) AAT-806 Target: 5′-CACCGAAGAGGCCAAGAAACAGATCAA-3′ (SEQ ID NO: 2396) 5′-CGAAGAGGCCAAGAAACAGAUCAac-3′ (SEQ ID NO: 2117) 3′-UGGCUUCUCCGGUUCUUUGUCUAGUUG-5′ (SEQ ID NO: 2257) AAT-807 Target: 5′-ACCGAAGAGGCCAAGAAACAGATCAAC-3′ (SEQ ID NO: 2397) 5′-GAAGAGGCCAAGAAACAGAUCAAcg-3′ (SEQ ID NO: 2118) 3′-GGCUUCUCCGGUUCUUUGUCUAGUUGC-5′ (SEQ ID NO: 2258) AAT-808 Target: 5′-CCGAAGAGGCCAAGAAACAGATCAACG-3′ (SEQ ID NO: 2398) 5′-AAGAGGCCAAGAAACAGAUCAACga-3′ (SEQ ID NO: 2119) 3′-GCUUCUCCGGUUCUUUGUCUAGUUGCU-5′ (SEQ ID NO: 2259) AAT-809 Target: 5′-CGAAGAGGCCAAGAAACAGATCAACGA-3′ (SEQ ID NO: 2399) 5′-AGAGGCCAAGAAACAGAUCAACGat-3′ (SEQ ID NO: 2120) 3′-CUUCUCCGGUUCUUUGUCUAGUUGCUA-5′ (SEQ ID NO: 2260) AAT-810 Target: 5′-GAAGAGGCCAAGAAACAGATCAACGAT-3′ (SEQ ID NO: 2400) 5′-GAGGCCAAGAAACAGAUCAACGAtt-3′ (SEQ ID NO: 2121) 3′-UUCUCCGGUUCUUUGUCUAGUUGCUAA-5′ (SEQ ID NO: 2261) AAT-811 Target: 5′-AAGAGGCCAAGAAACAGATCAACGATT-3′ (SEQ ID NO: 2401) 5′-AGGCCAAGAAACAGAUCAACGAUta-3′ (SEQ ID NO: 2122) 3′-UCUCCGGUUCUUUGUCUAGUUGCUAAU-5′ (SEQ ID NO: 2262) AAT-812 Target: 5′-AGAGGCCAAGAAACAGATCAACGATTA-3′ (SEQ ID NO: 2402) 5′-GGCCAAGAAACAGAUCAACGAUUac-3′ (SEQ ID NO: 2123) 3′-CUCCGGUUCUUUGUCUAGUUGCUAAUG-5′ (SEQ ID NO: 2263) AAT-813 Target: 5′-GAGGCCAAGAAACAGATCAACGATTAC-3′ (SEQ ID NO: 2403) 5′-UUUUGCUCUGGUGAAUUACAUCUtc-3′ (SEQ ID NO: 2124) 3′-CAAAAACGAGACCACUUAAUGUAGAAG-5′ (SEQ ID NO: 2264) AAT-900 Target: 5′-GTTTTTGCTCTGGTGAATTACATCTTC-3′ (SEQ ID NO: 2404) 5′-UUUGCUCUGGUGAAUUACAUCUUct-3′ (SEQ ID NO: 2125) 3′-AAAAACGAGACCACUUAAUGUAGAAGA-5′ (SEQ ID NO: 2265) AAT-901 Target: 5′-TTTTTGCTCTGGTGAATTACATCTTCT-3′ (SEQ ID NO: 2405) 5′-UUGCUCUGGUGAAUUACAUCUUCtt-3′ (SEQ ID NO: 2126) 3′-AAAACGAGACCACUUAAUGUAGAAGAA-5′ (SEQ ID NO: 2266) AAT-902 Target: 5′-TTTTGCTCTGGTGAATTACATCTTCTT-3′ (SEQ ID NO: 2406) 5′-UGCUCUGGUGAAUUACAUCUUCUtt-3′ (SEQ ID NO: 2127) 3′-AAACGAGACCACUUAAUGUAGAAGAAA-5′ (SEQ ID NO: 2267) AAT-903 Target: 5′-TTTGCTCTGGTGAATTACATCTTCTTT-3′ (SEQ ID NO: 2407) 5′-GCUCUGGUGAAUUACAUCUUCUUta-3′ (SEQ ID NO: 2128) 3′-AACGAGACCACUUAAUGUAGAAGAAAU-5′ (SEQ ID NO: 2268) AAT-904 Target: 5′-TTGCTCTGGTGAATTACATCTTCTTTA-3′ (SEQ ID NO: 2408) 5′-CUCUGGUGAAUUACAUCUUCUUUaa-3′ (SEQ ID NO: 2129) 3′-ACGAGACCACUUAAUGUAGAAGAAAUU-5′ (SEQ ID NO: 2269) AAT-905 Target: 5′-TGCTCTGGTGAATTACATCTTCTTTAA-3′ (SEQ ID NO: 2409) 5′-UCUGGUGAAUUACAUCUUCUUUAaa-3′ (SEQ ID NO: 2130) 3′-CGAGACCACUUAAUGUAGAAGAAAUUU-5′ (SEQ ID NO: 2270) AAT-906 Target: 5′-GCTCTGGTGAATTACATCTTCTTTAAA-3′ (SEQ ID NO: 2410) 5′-CUGGUGAAUUACAUCUUCUUUAAag-3′(SEQ ID NO: 2131) 3′-GAGACCACUUAAUGUAGAAGAAAUUUC-5′ (SEQ ID NO: 2271) AAT-907 Target: 5′-CTCTGGTGAATTACATCTTCTTTAAAG-3′ (SEQ ID NO: 2411) 5′-UGGUGAAUUACAUCUUCUUUAAAgg-3′ (SEQ ID NO: 2132) 3′-AGACCACUUAAUGUAGAAGAAAUUUCC-5′ (SEQ ID NO: 2272) AAT-908 Target: 5′-TCTGGTGAATTACATCTTCTTTAAAGG-3′ (SEQ ID NO: 2412) 5′-GGUGAAUUACAUCUUCUUUAAAGgc-3′ (SEQ ID NO: 2133) 3′-GACCACUUAAUGUAGAAGAAAUUUCCG-5′ (SEQ ID NO: 2273) AAT-909 Target: 5′-CTGGTGAATTACATCTTCTTTAAAGGC-3′ (SEQ ID NO: 2413) 5′-GUGAAUUACAUCUUCUUUAAAGGca-3′ (SEQ ID NO: 2134) 3′-ACCACUUAAUGUAGAAGAAAUUUCCGU-5′ (SEQ ID NO: 2274) AAT-910 Target: 5′-TGGTGAATTACATCTTCTTTAAAGGCA-3′ (SEQ ID NO: 2414) 5′-UGAAUUACAUCUUCUUUAAAGGCaa-3′ (SEQ ID NO: 2135) 3′-CCACUUAAUGUAGAAGAAAUUUCCGUU-5′ (SEQ ID NO: 2275) AAT-911 Target: 5′-GGTGAATTACATCTTCTTTAAAGGCAA-3′ (SEQ ID NO: 2415) 5′-GAAUUACAUCUUCUUUAAAGGCAaa-3′ (SEQ ID NO: 2136) 3′-CACUUAAUGUAGAAGAAAUUUCCGUUU-5′ (SEQ ID NO: 2276) AAT-912 Target: 5′-GTGAATTACATCTTCTTTAAAGGCAAA-3′ (SEQ ID NO: 2416) 5′-AAUUACAUCUUCUUUAAAGGCAAat-3′ (SEQ ID NO: 2137) 3′-ACUUAAUGUAGAAGAAAUUUCCGUUUA-5′ (SEQ ID NO: 2277) AAT-913 Target: 5′-TGAATTACATCTTCTTTAAAGGCAAAT-3′ (SEQ ID NO: 2417) 5′-AUUACAUCUUCUUUAAAGGCAAAtg-3′ (SEQ ID NO: 2138) 3′-CUUAAUGUAGAAGAAAUUUCCGUUUAC-5′ (SEQ ID NO: 2278) AAT-914 Target: 5′-GAATTACATCTTCTTTAAAGGCAAATG-3′ (SEQ ID NO: 2418) 5′-UUACAUCUUCUUUAAAGGCAAAUgg-3′ (SEQ ID NO: 2139) 3′-UUAAUGUAGAAGAAAUUUCCGUUUACC-5′ (SEQ ID NO: 2279) AAT-915 Target: 5′-AATTACATCTTCTTTAAAGGCAAATGG-3′ (SEQ ID NO: 2419) 5′-UACAUCUUCUUUAAAGGCAAAUGgg-3′ (SEQ ID NO: 2140) 3′-UAAUGUAGAAGAAAUUUCCGUUUACCC-5′ (SEQ ID NO: 2280) AAT-916 Target: 5′-ATTACATCTTCTTTAAAGGCAAATGGG-3′ (SEQ ID NO: 2420) 5′-ACAUCUUCUUUAAAGGCAAAUGGga-3′ (SEQ ID NO: 2141) 3′-AAUGUAGAAGAAAUUUCCGUUUACCCU-5′ (SEQ ID NO: 2281) AAT-917 Target: 5′-TTACATCTTCTTTAAAGGCAAATGGGA-3′ (SEQ ID NO: 2421) 5′-CAUCUUCUUUAAAGGCAAAUGGGag-3′ (SEQ ID NO: 2142) 3′-AUGUAGAAGAAAUUUCCGUUUACCCUC-5′ (SEQ ID NO: 2282) AAT-918 Target: 5′-TACATCTTCTTTAAAGGCAAATGGGAG-3′ (SEQ ID NO: 2422) 5′-UUCUUUAAAGGCAAAUGGGAGAGac-3′ (SEQ ID NO: 2143) 3′-AGAAGAAAUUUCCGUUUACCCUCUCUG-5′ (SEQ ID NO: 2283) AAT-922 Target: 5′-TCTTCTTTAAAGGCAAATGGGAGAGAC-3′ (SEQ ID NO: 2423) 5′-CUUUAAAGGCAAAUGGGAGAGACcc-3′ (SEQ ID NO: 2144) 3′-AAGAAAUUUCCGUUUACCCUCUCUGGG-5′ (SEQ ID NO: 2284) AAT-924 Target: 5′-TTCTTTAAAGGCAAATGGGAGAGACCC-3′ (SEQ ID NO: 2424) 5′-GCAAAUGGGAGAGACCCUUUGAAgt-3′ (SEQ ID NO: 2145) 3′-UCCGUUUACCCUCUCUGGGAAACUUCA-5′ (SEQ ID NO: 2285) AAT-932 Target: 5′-AGGCAAATGGGAGAGACCCTTTGAAGT-3′ (SEQ ID NO: 2425) 5′-CAAAUGGGAGAGACCCUUUGAAGtc-3′ (SEQ ID NO: 2146) 3′-CCGUUUACCCUCUCUGGGAAACUUCAG-5′ (SEQ ID NO: 2286) AAT-933 Target: 5′-GGCAAATGGGAGAGACCCTTTGAAGTC-3′ (SEQ ID NO: 2426) 5′-AAAUGGGAGAGACCCUUUGAAGUca-3′ (SEQ ID NO: 2147) 3′-CGUUUACCCUCUCUGGGAAACUUCAGU-5′ (SEQ ID NO: 2287) AAT-934 Target: 5′-GCAAATGGGAGAGACCCTTTGAAGTCA-3′ (SEQ ID NO: 2427) 5′-AAUGGGAGAGACCCUUUGAAGUCaa-3′ (SEQ ID NO: 2148) 3′-GUUUACCCUCUCUGGGAAACUUCAGUU-5′ (SEQ ID NO: 2288) AAT-935 Target: 5′-CAAATGGGAGAGACCCTTTGAAGTCAA-3′ (SEQ ID NO: 2428) 5′-UGUCCAGCUGGGUGCUGCUGAUGaa-3′ (SEQ ID NO: 2149) 3′-CGACAGGUCGACCCACGACGACUACUU-5′ (SEQ ID NO: 2289) AAT-1061 Target: 5′-GCTGTCCAGCTGGGTGCTGCTGATGAA-3′ (SEQ ID NO: 2429) 5′-GUCCAGCUGGGUGCUGCUGAUGAaa-3′ (SEQ ID NO: 2150) 3′-GACAGGUCGACCCACGACGACUACUUU-5′ (SEQ ID NO: 2290) AAT-1062 Target: 5′-CTGTCCAGCTGGGTGCTGCTGATGAAA-3′ (SEQ ID NO: 2430) 5′-UCCAGCUGGGUGCUGCUGAUGAAat-3′ (SEQ ID NO: 2151) 3′-ACAGGUCGACCCACGACGACUACUUUA-5′ (SEQ ID NO: 2291) AAT-1063 Target: 5′-TGTCCAGCTGGGTGCTGCTGATGAAAT-3′ (SEQ ID NO: 2431) 5′-CCAGCUGGGUGCUGCUGAUGAAAta-3′ (SEQ ID NO: 2152) 3′-CAGGUCGACCCACGACGACUACUUUAU-5′ (SEQ ID NO: 2292) AAT-1064 Target: 5′-GTCCAGCTGGGTGCTGCTGATGAAATA-3′ (SEQ ID NO: 2432) 5′-CAGCUGGGUGCUGCUGAUGAAAUac-3′ (SEQ ID NO: 2153) 3′-AGGUCGACCCACGACGACUACUUUAUG-5′ (SEQ ID NO: 2293) AAT-1065 Target: 5′-TCCAGCTGGGTGCTGCTGATGAAATAC-3′ (SEQ ID NO: 2433) 5′-AGCUGGGUGCUGCUGAUGAAAUAcc-3′ (SEQ ID NO: 2154) 3′-GGUCGACCCACGACGACUACUUUAUGG-5′ (SEQ ID NO: 2294) AAT-1066 Target: 5′-CCAGCTGGGTGCTGCTGATGAAATACC-3′ (SEQ ID NO: 2434) 5′-GCUGGGUGCUGCUGAUGAAAUACct-3′ (SEQ ID NO: 2155) 3′-GUCGACCCACGACGACUACUUUAUGGA-5′ (SEQ ID NO: 2295) AAT-1067 Target: 5′-CAGCTGGGTGCTGCTGATGAAATACCT-3′ (SEQ ID NO: 2435) 5′-CUGGGUGCUGCUGAUGAAAUACCtg-3′ (SEQ ID NO: 2156) 3′-UCGACCCACGACGACUACUUUAUGGAC-5′ (SEQ ID NO: 2296) AAT-1068 Target: 5′-AGCTGGGTGCTGCTGATGAAATACCTG-3′ (SEQ ID NO: 2436) 5′-UGGGUGCUGCUGAUGAAAUACCUgg-3′ (SEQ ID NO: 2157) 3′-CGACCCACGACGACUACUUUAUGGACC-5′ (SEQ ID NO: 2297) AAT-1069 Target: 5′-GCTGGGTGCTGCTGATGAAATACCTGG-3′ (SEQ ID NO: 2437) 5′-GGGUGCUGCUGAUGAAAUACCUGgg-3′ (SEQ ID NO: 2158) 3′-GACCCACGACGACUACUUUAUGGACCC-5′ (SEQ ID NO: 2298) AAT-1070 Target: 5′-CTGGGTGCTGCTGATGAAATACCTGGG-3′ (SEQ ID NO: 2438) 5′-GUGCUGCUGAUGAAAUACCUGGGca-3′ (SEQ ID NO: 2159) 3′-CCCACGACGACUACUUUAUGGACCCGU-5′ (SEQ ID NO: 2299) AAT-1072 Target: 5′-GGGTGCTGCTGATGAAATACCTGGGCA-3′ (SEQ ID NO: 2439) 5′-UGCUGCUGAUGAAAUACCUGGGCaa-3′ (SEQ ID NO: 2160) 3′-CCACGACGACUACUUUAUGGACCCGUU-5′ (SEQ ID NO: 2300) AAT-1073 Target: 5′-GGTGCTGCTGATGAAATACCTGGGCAA-3′ (SEQ ID NO: 2440) 5′-GCUGCUGAUGAAAUACCUGGGCAat-3′ (SEQ ID NO: 2161) 3′-CACGACGACUACUUUAUGGACCCGUUA-5′ (SEQ ID NO: 2301) AAT-1074 Target: 5′-GTGCTGCTGATGAAATACCTGGGCAAT-3′ (SEQ ID NO: 2441) 5′-CUGCUGAUGAAAUACCUGGGCAAtg-3′ (SEQ ID NO: 2162) 3′-ACGACGACUACUUUAUGGACCCGUUAC-5′ (SEQ ID NO: 2302) AAT-1075 Target: 5′-TGCTGCTGATGAAATACCTGGGCAATG-3′ (SEQ ID NO: 2442) 5′-UGCUGAUGAAAUACCUGGGCAAUgc-3′ (SEQ ID NO: 2163) 3′-CGACGACUACUUUAUGGACCCGUUACG-5′ (SEQ ID NO: 2303) AAT-1076 Target: 5′-GCTGCTGATGAAATACCTGGGCAATGC-3′ (SEQ ID NO: 2443) 5′-GCUGAUGAAAUACCUGGGCAAUGcc-3′ (SEQ ID NO: 2164) 3′-GACGACUACUUUAUGGACCCGUUACGG-5′ (SEQ ID NO: 2304) AAT-1077 Target: 5′-CTGCTGATGAAATACCTGGGCAATGCC-3′ (SEQ ID NO: 2444) 5′-CUGAUGAAAUACCUGGGCAAUGCca-3′ (SEQ ID NO: 2165) 3′-ACGACUACUUUAUGGACCCGUUACGGU-5′ (SEQ ID NO: 2305) AAT-1078 Target: 5′-TGCTGATGAAATACCTGGGCAATGCCA-3′ (SEQ ID NO: 2445) 5′-UGAUGAAAUACCUGGGCAAUGCCac-3′ (SEQ ID NO: 2166) 3′-CGACUACUUUAUGGACCCGUUACGGUG-5′ (SEQ ID NO: 2306) AAT-1079 Target: 5′-GCTGATGAAATACCTGGGCAATGCCAC-3′ (SEQ ID NO: 2446) 5′-GAUGAAAUACCUGGGCAAUGCCAcc-3′ (SEQ ID NO: 2167) 3′-GACUACUUUAUGGACCCGUUACGGUGG-5′ (SEQ ID NO: 2307) AAT-1080 Target: 5′-CTGATGAAATACCTGGGCAATGCCACC-3′ (SEQ ID NO: 2447) 5′-AUGAAAUACCUGGGCAAUGCCACcg-3′ (SEQ ID NO: 2168) 3′-ACUACUUUAUGGACCCGUUACGGUGGC-5′ (SEQ ID NO: 2308) AAT-1081 Target: 5′-TGATGAAATACCTGGGCAATGCCACCG-3′ (SEQ ID NO: 2448) 5′-GAAAUACCUGGGCAAUGCCACCGcc-3′ (SEQ ID NO: 2169) 3′-UACUUUAUGGACCCGUUACGGUGGCGG-5′ (SEQ ID NO: 2309) AAT-1083 Target: 5′-ATGAAATACCTGGGCAATGCCACCGCC-3′ (SEQ ID NO: 2449) 5′-CAGCACCUGGAAAAUGAACUCACcc-3′ (SEQ ID NO: 2170) 3′-AUGUCGUGGACCUUUUACUUGAGUGGG-5′ (SEQ ID NO: 2310) AAT-1138 Target: 5′-TACAGCACCTGGAAAATGAACTCACCC-3′ (SEQ ID NO: 2450) 5′-CUGGAAAAUGAACUCACCCACGAta-3′ (SEQ ID NO: 2171) 3′-UGGACCUUUUACUUGAGUGGGUGCUAU-5′ (SEQ ID NO: 2311) AAT-1144 Target: 5′-ACCTGGAAAATGAACTCACCCACGATA-3′ (SEQ ID NO: 2451) 5′-UGGAAAAUGAACUCACCCACGAUat-3′ (SEQ ID NO: 2172) 3′-GGACCUUUUACUUGAGUGGGUGCUAUA-5′ (SEQ ID NO: 2312) AAT-1145 Target: 5′-CCTGGAAAATGAACTCACCCACGATAT-3′ (SEQ ID NO: 2452) 5′-GAUAUCAUCACCAAGUUCCUGGAaa-3′ (SEQ ID NO: 2173) 3′-UGCUAUAGUAGUGGUUCAAGGACCUUU-5′ (SEQ ID NO: 2313) AAT-1165 Target: 5′-ACGATATCATCACCAAGTTCCTGGAAA-3′ (SEQ ID NO: 2453) 5′-CAAGUUCCUGGAAAAUGAAGACAga-3′ (SEQ ID NO: 2174) 3′-UGGUUCAAGGACCUUUUACUUCUGUCU-5′ (SEQ ID NO: 2314) AAT-1176 Target: 5′-ACCAAGTTCCTGGAAAATGAAGACAGA-3′ (SEQ ID NO: 2454) 5′-CCAUUACUGGAACCUAUGAUCUGaa-3′ (SEQ ID NO: 2175) 3′-CAGGUAAUGACCUUGGAUACUAGACUU-5′ (SEQ ID NO: 2315) AAT-1232 Target: 5′-GTCCATTACTGGAACCTATGATCTGAA-3′ (SEQ ID NO: 2455) 5′-CAUUACUGGAACCUAUGAUCUGAag-3′ (SEQ ID NO: 2176) 3′-AGGUAAUGACCUUGGAUACUAGACUUC-5′ (SEQ ID NO: 2316) AAT-1233 Target: 5′-TCCATTACTGGAACCTATGATCTGAAG-3′ (SEQ ID NO: 2456) 5′-AUUACUGGAACCUAUGAUCUGAAga-3′ (SEQ ID NO: 2177) 3′-GGUAAUGACCUUGGAUACUAGACUUCU-5′ (SEQ ID NO: 2317) AAT-1234 Target: 5′-CCATTACTGGAACCTATGATCTGAAGA-3′ (SEQ ID NO: 2457) 5′-UUACUGGAACCUAUGAUCUGAAGag-3′ (SEQ ID NO: 2178) 3′-GUAAUGACCUUGGAUACUAGACUUCUC-5′ (SEQ ID NO: 2318) AAT-1235 Target: 5′-CATTACTGGAACCTATGATCTGAAGAG-3′ (SEQ ID NO: 2458) 5′-UACUGGAACCUAUGAUCUGAAGAgc-3′ (SEQ ID NO: 2179) 3′-UAAUGACCUUGGAUACUAGACUUCUCG-5′ (SEQ ID NO: 2319) AAT-1236 Target: 5′-ATTACTGGAACCTATGATCTGAAGAGC-3′ (SEQ ID NO: 2459) 5′-ACUGGAACCUAUGAUCUGAAGAGcg-3′ (SEQ ID NO: 2180) 3′-AAUGACCUUGGAUACUAGACUUCUCGC-5′ (SEQ ID NO: 2320) AAT-1237 Target: 5′-TTACTGGAACCTATGATCTGAAGAGCG-3′ (SEQ ID NO: 2460) 5′-CUGGAACCUAUGAUCUGAAGAGCgt-3′ (SEQ ID NO: 2181) 3′-AUGACCUUGGAUACUAGACUUCUCGCA-5′ (SEQ ID NO: 2321) AAT-1238 Target: 5′-TACTGGAACCTATGATCTGAAGAGCGT-3′ (SEQ ID NO: 2461) 5′-UGGAACCUAUGAUCUGAAGAGCGtc-3′ (SEQ ID NO: 2182) 3′-UGACCUUGGAUACUAGACUUCUCGCAG-5′ (SEQ ID NO: 2322) AAT-1239 Target: 5′-ACTGGAACCTATGATCTGAAGAGCGTC-3′ (SEQ ID NO: 2462) 5′-GGAACCUAUGAUCUGAAGAGCGUcc-3′ (SEQ ID NO: 2183) 3′-GACCUUGGAUACUAGACUUCUCGCAGG-5′ (SEQ ID NO: 2323) AAT-1240 Target: 5′-CTGGAACCTATGATCTGAAGAGCGTCC-3′ (SEQ ID NO: 2463) 5′-AUCACUAAGGUCUUCAGCAAUGGgg-3′ (SEQ ID NO: 2184) 3′-CGUAGUGAUUCCAGAAGUCGUUACCCC-5′ (SEQ ID NO: 2324) AAT-1279 Target: 5′-GCATCACTAAGGTCTTCAGCAATGGGG-3′ (SEQ ID NO: 2464) 5′-UCACUAAGGUCUUCAGCAAUGGGgc-3′ (SEQ ID NO: 2185) 3′-GUAGUGAUUCCAGAAGUCGUUACCCCG-5′ (SEQ ID NO: 2325) AAT-1280 Target: 5′-CATCACTAAGGTCTTCAGCAATGGGGC-3′ (SEQ ID NO: 2465) 5′-CACUAAGGUCUUCAGCAAUGGGGct-3′ (SEQ ID NO: 2186) 3′-UAGUGAUUCCAGAAGUCGUUACCCCGA-5′ (SEQ ID NO: 2326) AAT-1281 Target: 5′-ATCACTAAGGTCTTCAGCAATGGGGCT-3′ (SEQ ID NO: 2466) 5′-CUAAGGUCUUCAGCAAUGGGGCUga-3′ (SEQ ID NO: 2187) 3′-GUGAUUCCAGAAGUCGUUACCCCGACU-5′ (SEQ ID NO: 2327) AAT-1283 Target: 5′-CACTAAGGTCTTCAGCAATGGGGCTGA-3′ (SEQ ID NO: 2467) 5′-UAAGGUCUUCAGCAAUGGGGCUGac-3′ (SEQ ID NO: 2188) 3′-UGAUUCCAGAAGUCGUUACCCCGACUG-5′ (SEQ ID NO: 2328) AAT-1284 Target: 5′-ACTAAGGTCTTCAGCAATGGGGCTGAC-3′ (SEQ ID NO: 2468) 5′-UGAAGCUCUCCAAGGCCGUGCAUaa-3′ (SEQ ID NO: 2189) 3′-GGACUUCGAGAGGUUCCGGCACGUAUU-5′ (SEQ ID NO: 2329) AAT-1337 Target: 5′-CCTGAAGCTCTCCAAGGCCGTGCATAA-3′ (SEQ ID NO: 2469) 5′-GAAGCUCUCCAAGGCCGUGCAUAag-3′ (SEQ ID NO: 2190) 3′-GACUUCGAGAGGUUCCGGCACGUAUUC-5′ (SEQ ID NO: 2330) AAT-1338 Target: 5′-CTGAAGCTCTCCAAGGCCGTGCATAAG-3′ (SEQ ID NO: 2470) 5′-AAGCUCUCCAAGGCCGUGCAUAAgg-3′ (SEQ ID NO: 2191) 3′-ACUUCGAGAGGUUCCGGCACGUAUUCC-5′ (SEQ ID NO: 2331) AAT-1339 Target: 5′-TGAAGCTCTCCAAGGCCGTGCATAAGG-3′ (SEQ ID NO: 2471) 5′-CCGAGGUCAAGUUCAACAAACCCtt-3′ (SEQ ID NO: 2192) 3′-GGGGCUCCAGUUCAAGUUGUUUGGGAA-5′ (SEQ ID NO: 2332) AAT-1442 Target: 5′-CCCCGAGGTCAAGTTCAACAAACCCTT-3′ (SEQ ID NO: 2472) 5′-CGAGGUCAAGUUCAACAAACCCUtt-3′ (SEQ ID NO: 2193) 3′-GGGCUCCAGUUCAAGUUGUUUGGGAAA-5′ (SEQ ID NO: 2333) AAT-1443 Target: 5′-CCCGAGGTCAAGTTCAACAAACCCTTT-3′ (SEQ ID NO: 2473) 5′-GAGGUCAAGUUCAACAAACCCUUtg-3′ (SEQ ID NO: 2194) 3′-GGCUCCAGUUCAAGUUGUUUGGGAAAC-5′ (SEQ ID NO: 2334) AAT-1444 Target: 5′-CCGAGGTCAAGTTCAACAAACCCTTTG-3′ (SEQ ID NO: 2474) 5′-AGGUCAAGUUCAACAAACCCUUUgt-3′ (SEQ ID NO: 2195) 3′-GCUCCAGUUCAAGUUGUUUGGGAAACA-5′ (SEQ ID NO: 2335) AAT-1445 Target: 5′-CGAGGTCAAGTTCAACAAACCCTTTGT-3′ (SEQ ID NO: 2475) 5′-GGUCAAGUUCAACAAACCCUUUGtc-3′ (SEQ ID NO: 2196) 3′-CUCCAGUUCAAGUUGUUUGGGAAACAG-5′ (SEQ ID NO: 2336) AAT-1446 Target: 5′-GAGGTCAAGTTCAACAAACCCTTTGTC-3′ (SEQ ID NO: 2476) 5′-GUCAAGUUCAACAAACCCUUUGUct-3′ (SEQ ID NO: 2197) 3′-UCCAGUUCAAGUUGUUUGGGAAACAGA-5′ (SEQ ID NO: 2337) AAT-1447 Target: 5′-AGGTCAAGTTCAACAAACCCTTTGTCT-3′ (SEQ ID NO: 2477) 5′-UCAAGUUCAACAAACCCUUUGUCtt-3′ (SEQ ID NO: 2198) 3′-CCAGUUCAAGUUGUUUGGGAAACAGAA-5′ (SEQ ID NO: 2338) AAT-1448 Target: 5′-GGTCAAGTTCAACAAACCCTTTGTCTT-3′ (SEQ ID NO: 2478) 5′-CAAGUUCAACAAACCCUUUGUCUtc-3′ (SEQ ID NO: 2199) 3′-CAGUUCAAGUUGUUUGGGAAACAGAAG-5′ (SEQ ID NO: 2339) AAT-1449 Target: 5′-GTCAAGTTCAACAAACCCTTTGTCTTC-3′ (SEQ ID NO: 2479) 5′-AAGUUCAACAAACCCUUUGUCUUct-3′ (SEQ ID NO: 2200) 3′-AGUUCAAGUUGUUUGGGAAACAGAAGA-5′ (SEQ ID NO: 2340) AAT-1450 Target: 5′-TCAAGTTCAACAAACCCTTTGTCTTCT-3′ (SEQ ID NO: 2480) 5′-AGUUCAACAAACCCUUUGUCUUCtt-3′ (SEQ ID NO: 2201) 3′-GUUCAAGUUGUUUGGGAAACAGAAGAA-5′(SEQ ID NO: 2341) AAT-1451 Target: 5′-CAAGTTCAACAAACCCTTTGTCTTCTT-3′ (SEQ ID NO: 2481) 

TABLE 13 Further Human Anti-a-1 antitrypsin DsiRNAs, Unmodified Duplexes  (Asymmetrics)  5′-CCAGGGAGAUGCUGCCCAGAAGACA-3′ (SEQ ID NO: 2482) 3′-GGGGUCCCUCUACGACGGGUCUUCUGU-5′ (SEQ ID NO: 2202) AAT-366  Target: 5′-CCCCAGGGAGATGCTGCCCAGAAGACA-3′ (SEQ ID NO: 2342) 5′-CAGGGAGAUGCUGCCCAGAAGACAG-3′ (SEQ ID NO: 2483) 3′-GGGUCCCUCUACGACGGGUCUUCUGUC-5′ (SEQ ID NO: 2203) AAT-367  Target: 5′-CCCAGGGAGATGCTGCCCAGAAGACAG-3′ (SEQ ID NO: 2343) 5′-AGGGAGAUGCUGCCCAGAAGACAGA-3′ (SEQ ID NO: 2484) 3′-GGUCCCUCUACGACGGGUCUUCUGUCU-5′ (SEQ ID NO: 2204) AAT-368  Target: 5′-CCAGGGAGATGCTGCCCAGAAGACAGA-3′ (SEQ ID NO: 2344) 5′-GGGAGAUGCUGCCCAGAAGACAGAU-3′ (SEQ ID NO: 2485) 3′-GUCCCUCUACGACGGGUCUUCUGUCUA-5′ (SEQ ID NO: 2205) AAT-369  Target: 5′-CAGGGAGATGCTGCCCAGAAGACAGAT-3′ (SEQ ID NO: 2345) 5′-GGAGAUGCUGCCCAGAAGACAGAUA-3′ (SEQ ID NO: 2486) 3′-UCCCUCUACGACGGGUCUUCUGUCUAU-5′ (SEQ ID NO: 2206) AAT-370  Target: 5′-AGGGAGATGCTGCCCAGAAGACAGATA-3′ (SEQ ID NO: 2346) 5′-GAGAUGCUGCCCAGAAGACAGAUAC-3′ (SEQ ID NO: 2487) 3′-CCCUCUACGACGGGUCUUCUGUCUAUG-5′ (SEQ ID NO: 2207) AAT-371  Target: 5′-GGGAGATGCTGCCCAGAAGACAGATAC-3′ (SEQ ID NO: 2347) 5′-GAUACAUCCCACCAUGAUCAGGAUC-3′ (SEQ ID NO: 2488) 3′-GUCUAUGUAGGGUGGUACUAGUCCUAG-5′ (SEQ ID NO: 2208) AAT-391  Target: 5′-CAGATACATCCCACCATGATCAGGATC-3′ (SEQ ID NO: 2348) 5′-AUACAUCCCACCAUGAUCAGGAUCA-3′ (SEQ ID NO: 2489) 3′-UCUAUGUAGGGUGGUACUAGUCCUAGU-5′ (SEQ ID NO: 2209) AAT-392  Target: 5′-AGATACATCCCACCATGATCAGGATCA-3′ (SEQ ID NO: 2349) 5′-UACAUCCCACCAUGAUCAGGAUCAC-3′ (SEQ ID NO: 2490) 3′-CUAUGUAGGGUGGUACUAGUCCUAGUG-5′ (SEQ ID NO: 2210) AAT-393  Target: 5′-GATACATCCCACCATGATCAGGATCAC-3′ (SEQ ID NO: 2350) 5′-ACAUCCCACCAUGAUCAGGAUCACC-3′ (SEQ ID NO: 2491) 3′-UAUGUAGGGUGGUACUAGUCCUAGUGG-5′ (SEQ ID NO: 2211) AAT-394  Target: 5′-ATACATCCCACCATGATCAGGATCACC-3′ (SEQ ID NO: 2351) 5′-ACCAGUCCAACAGCACCAAUAUCUU-3′ (SEQ ID NO: 2492) 3′-UGUGGUCAGGUUGUCGUGGUUAUAGAA-5′ (SEQ ID NO: 2212) AAT-485  Target: 5′-ACACCAGTCCAACAGCACCAATATCTT-3′ (SEQ ID NO: 2352) 5′-CCAGUCCAACAGCACCAAUAUCUUC-3′ (SEQ ID NO: 2493) 3′-GUGGUCAGGUUGUCGUGGUUAUAGAAG-5′ (SEQ ID NO: 2213) AAT-486  Target: 5′-CACCAGTCCAACAGCACCAATATCTTC-3′ (SEQ ID NO: 2353) 5′-CAGUCCAACAGCACCAAUAUCUUCU-3′ (SEQ ID NO: 2494) 3′-UGGUCAGGUUGUCGUGGUUAUAGAAGA-5′ (SEQ ID NO: 2214) AAT-487  Target: 5′-ACCAGTCCAACAGCACCAATATCTTCT-3′ (SEQ ID NO: 2354) 5′-AGUCCAACAGCACCAAUAUCUUCUU-3′ (SEQ ID NO: 2495) 3′-GGUCAGGUUGUCGUGGUUAUAGAAGAA-5′ (SEQ ID NO: 2215) AAT-488  Target: 5′-CCAGTCCAACAGCACCAATATCTTCTT-3′ (SEQ ID NO: 2355) 5′-GUCCAACAGCACCAAUAUCUUCUUC-3′ (SEQ ID NO: 2496) 3′-GUCAGGUUGUCGUGGUUAUAGAAGAAG-5′ (SEQ ID NO: 2216) AAT-489  Target: 5′-CAGTCCAACAGCACCAATATCTTCTTC-3′ (SEQ ID NO: 2356) 5′-UCCAACAGCACCAAUAUCUUCUUCU-3′ (SEQ ID NO: 2497) 3′-UCAGGUUGUCGUGGUUAUAGAAGAAGA-5′ (SEQ ID NO: 2217) AAT-490  Target: 5′-AGTCCAACAGCACCAATATCTTCTTCT-3′ (SEQ ID NO: 2357) 5′-CCAACAGCACCAAUAUCUUCUUCUC-3′ (SEQ ID NO: 2498) 3′-CAGGUUGUCGUGGUUAUAGAAGAAGAG-5′ (SEQ ID NO: 2218) AAT-491  Target: 5′-GTCCAACAGCACCAATATCTTCTTCTC-3′ (SEQ ID NO: 2358) 5′-CAACAGCACCAAUAUCUUCUUCUCC-3′ (SEQ ID NO: 2499) 3′-AGGUUGUCGUGGUUAUAGAAGAAGAGG-5′ (SEQ ID NO: 2219) AAT-492  Target: 5′-TCCAACAGCACCAATATCTTCTTCTCC-3′ (SEQ ID NO: 2359) 5′-AACAGCACCAAUAUCUUCUUCUCCC-3′ (SEQ ID NO: 2500) 3′-GGUUGUCGUGGUUAUAGAAGAAGAGGG-5′ (SEQ ID NO: 2220) AAT-493  Target: 5′-CCAACAGCACCAATATCTTCTTCTCCC-3′ (SEQ ID NO: 2360) 5′-ACAGCACCAAUAUCUUCUUCUCCCC-3′ (SEQ ID NO: 2501) 3′-GUUGUCGUGGUUAUAGAAGAAGAGGGG-5′ (SEQ ID NO: 2221) AAT-494  Target: 5′-CAACAGCACCAATATCTTCTTCTCCCC-3′ (SEQ ID NO: 2361) 5′-CAGCACCAAUAUCUUCUUCUCCCCA-3′ (SEQ ID NO: 2502) 3′-UUGUCGUGGUUAUAGAAGAAGAGGGGU-5′ (SEQ ID NO: 2222) AAT-495  Target: 5′-AACAGCACCAATATCTTCTTCTCCCCA-3′ (SEQ ID NO: 2362) 5′-AGCACCAAUAUCUUCUUCUCCCCAG-3′ (SEQ ID NO: 2503) 3′-UGUCGUGGUUAUAGAAGAAGAGGGGUC-5′ (SEQ ID NO: 2223) AAT-496  Target: 5′-ACAGCACCAATATCTTCTTCTCCCCAG-3′ (SEQ ID NO: 2363) 5′-GCACCAAUAUCUUCUUCUCCCCAGU-3′ (SEQ ID NO: 2504) 3′-GUCGUGGUUAUAGAAGAAGAGGGGUCA-5′ (SEQ ID NO: 2224) AAT-497  Target: 5′-CAGCACCAATATCTTCTTCTCCCCAGT-3′ (SEQ ID NO: 2364) 5′-CACCAAUAUCUUCUUCUCCCCAGUG-3′ (SEQ ID NO: 2505) 3′-UCGUGGUUAUAGAAGAAGAGGGGUCAC-5′ (SEQ ID NO: 2225) AAT-498  Target: 5′-AGCACCAATATCTTCTTCTCCCCAGTG-3′ (SEQ ID NO: 2365) 5′-ACCAAUAUCUUCUUCUCCCCAGUGA-3′ (SEQ ID NO: 2506) 3′-CGUGGUUAUAGAAGAAGAGGGGUCACU-5′ (SEQ ID NO: 2226) AAT-499  Target: 5′-GCACCAATATCTTCTTCTCCCCAGTGA-3′ (SEQ ID NO: 2366) 5′-CCCAGUGAGCAUCGCUACAGCCUUU-3′ (SEQ ID NO: 2507) 3′-AGGGGUCACUCGUAGCGAUGUCGGAAA-5′ (SEQ ID NO: 2227) AAT-516  Target: 5′-TCCCCAGTGAGCATCGCTACAGCCTTT-3′ (SEQ ID NO: 2367) 5′-CCAGUGAGCAUCGCUACAGCCUUUG-3′ (SEQ ID NO: 2508) 3′-GGGGUCACUCGUAGCGAUGUCGGAAAC-5′ (SEQ ID NO: 2228) AAT-517  Target: 5′-CCCCAGTGAGCATCGCTACAGCCTTTG-3′ (SEQ ID NO: 2368) 5′-CAGUGAGCAUCGCUACAGCCUUUGC-3′ (SEQ ID NO: 2509) 3′-GGGUCACUCGUAGCGAUGUCGGAAACG-5′ (SEQ ID NO: 2229) AAT-518  Target: 5′-CCCAGTGAGCATCGCTACAGCCTTTGC-3′ (SEQ ID NO: 2369) 5′-AGUGAGCAUCGCUACAGCCUUUGCA-3′ (SEQ ID NO: 2510) 3′-GGUCACUCGUAGCGAUGUCGGAAACGU-5′ (SEQ ID NO: 2230) AAT-519  Target: 5′-CCAGTGAGCATCGCTACAGCCTTTGCA-3′ (SEQ ID NO: 2370) 5′-GUGAGCAUCGCUACAGCCUUUGCAA-3′ (SEQ ID NO: 2511) 3′-GUCACUCGUAGCGAUGUCGGAAACGUU-5′ (SEQ ID NO: 2231) AAT-520  Target: 5′-CAGTGAGCATCGCTACAGCCTTTGCAA-3′ (SEQ ID NO: 2371) 5′-UGAGCAUCGCUACAGCCUUUGCAAU-3′ (SEQ ID NO: 2512) 3′-UCACUCGUAGCGAUGUCGGAAACGUUA-5′ (SEQ ID NO: 2232) AAT-521  Target: 5′-AGTGAGCATCGCTACAGCCTTTGCAAT-3′ (SEQ ID NO: 2372) 5′-GAGCAUCGCUACAGCCUUUGCAAUG-3′ (SEQ ID NO: 2513) 3′-CACUCGUAGCGAUGUCGGAAACGUUAC-5′ (SEQ ID NO: 2233) AAT-522  Target: 5′-GTGAGCATCGCTACAGCCTTTGCAATG-3′ (SEQ ID NO: 2373) 5′-AGCAUCGCUACAGCCUUUGCAAUGC-3′ (SEQ ID NO: 2514) 3′-ACUCGUAGCGAUGUCGGAAACGUUACG-5′ (SEQ ID NO: 2234) AAT-523  Target: 5′-TGAGCATCGCTACAGCCTTTGCAATGC-3′ (SEQ ID NO: 2374) 5′-GCAUCGCUACAGCCUUUGCAAUGCU-3′ (SEQ ID NO: 2515) 3′-CUCGUAGCGAUGUCGGAAACGUUACGA-5′ (SEQ ID NO: 2235) AAT-524  Target: 5′-GAGCATCGCTACAGCCTTTGCAATGCT-3′ (SEQ ID NO: 2375) 5′-CAUCGCUACAGCCUUUGCAAUGCUC-3′ (SEQ ID NO: 2516) 3′-UCGUAGCGAUGUCGGAAACGUUACGAG-5′ (SEQ ID NO: 2236) AAT-525  Target: 5′-AGCATCGCTACAGCCTTTGCAATGCTC-3′ (SEQ ID NO: 2376) 5′-AUCGCUACAGCCUUUGCAAUGCUCU-3′ (SEQ ID NO: 2517) 3′-CGUAGCGAUGUCGGAAACGUUACGAGA-5′ (SEQ ID NO: 2237) AAT-526  Target: 5′-GCATCGCTACAGCCTTTGCAATGCTCT-3′ (SEQ ID NO: 2377) 5′-UCGCUACAGCCUUUGCAAUGCUCUC-3′ (SEQ ID NO: 2518) 3′-GUAGCGAUGUCGGAAACGUUACGAGAG-5′ (SEQ ID NO: 2238) AAT-527  Target: 5′-CATCGCTACAGCCTTTGCAATGCTCTC-3′ (SEQ ID NO: 2378) 5′-CGCUACAGCCUUUGCAAUGCUCUCC-3′ (SEQ ID NO: 2519) 3′-UAGCGAUGUCGGAAACGUUACGAGAGG-5′ (SEQ ID NO: 2239) AAT-528  Target: 5′-ATCGCTACAGCCTTTGCAATGCTCTCC-3′ (SEQ ID NO: 2379) 5′-GCUACAGCCUUUGCAAUGCUCUCCC-3′ (SEQ ID NO: 2520) 3′-AGCGAUGUCGGAAACGUUACGAGAGGG-5′ (SEQ ID NO: 2240) AAT-529  Target: 5′-TCGCTACAGCCTTTGCAATGCTCTCCC-3′ (SEQ ID NO: 2380) 5′-CUACAGCCUUUGCAAUGCUCUCCCU-3′ (SEQ ID NO: 2521) 3′-GCGAUGUCGGAAACGUUACGAGAGGGA-5′ (SEQ ID NO: 2241) AAT-530  Target: 5′-CGCTACAGCCTTTGCAATGCTCTCCCT-3′ (SEQ ID NO: 2381) 5′-UACAGCCUUUGCAAUGCUCUCCCUG-3′ (SEQ ID NO: 2522) 3′-CGAUGUCGGAAACGUUACGAGAGGGAC-5′ (SEQ ID NO: 2242) AAT-531  Target: 5′-GCTACAGCCTTTGCAATGCTCTCCCTG-3′ (SEQ ID NO: 2382) 5′-CCUGGGGACCAAGGCUGACACUCAC-3′ (SEQ ID NO: 2523) 3′-AGGGACCCCUGGUUCCGACUGUGAGUG-5′ (SEQ ID NO: 2243) AAT-552  Target: 5′-TCCCTGGGGACCAAGGCTGACACTCAC-3′ (SEQ ID NO: 2383) 5′-GGGACCAAGGCUGACACUCACGAUG-3′ (SEQ ID NO: 2524) 3′-ACCCCUGGUUCCGACUGUGAGUGCUAC-5′ (SEQ ID NO: 2244) AAT-556  Target: 5′-TGGGGACCAAGGCTGACACTCACGATG-3′ (SEQ ID NO: 2384) 5′-GGACCAAGGCUGACACUCACGAUGA-3′ (SEQ ID NO: 2525) 3′-CCCCUGGUUCCGACUGUGAGUGCUACU-5′ (SEQ ID NO: 2245) AAT-557  Target: 5′-GGGGACCAAGGCTGACACTCACGATGA-3′ (SEQ ID NO: 2385) 5′-GACCAAGGCUGACACUCACGAUGAA-3′ (SEQ ID NO: 2526) 3′-CCCUGGUUCCGACUGUGAGUGCUACUU-5′ (SEQ ID NO: 2246) AAT-558  Target: 5′-GGGACCAAGGCTGACACTCACGATGAA-3′ (SEQ ID NO: 2386) 5′-UGAAAUCCUGGAGGGCCUGAAUUUC-3′ (SEQ ID NO: 2527) 3′-CUACUUUAGGACCUCCCGGACUUAAAG-5′ (SEQ ID NO: 2247) AAT-579  Target: 5′-GATGAAATCCTGGAGGGCCTGAATTTC-3′ (SEQ ID NO: 2387) 5′-GAAAUCCUGGAGGGCCUGAAUUUCA-3′ (SEQ ID NO: 2528) 3′-UACUUUAGGACCUCCCGGACUUAAAGU-5′ (SEQ ID NO: 2248) AAT-580  Target: 5′-ATGAAATCCTGGAGGGCCTGAATTTCA-3′ (SEQ ID NO: 2388) 5′-UCCAUGAAGGCUUCCAGGAACUCCU-3′ (SEQ ID NO: 2529) 3′-CUAGGUACUUCCGAAGGUCCUUGAGGA-5′ (SEQ ID NO: 2249) AAT-632  Target: 5′-GATCCATGAAGGCTTCCAGGAACTCCT-3′ (SEQ ID NO: 2389) 5′-CCAUGAAGGCUUCCAGGAACUCCUC-3′ (SEQ ID NO: 2530) 3′-UAGGUACUUCCGAAGGUCCUUGAGGAG-5′ (SEQ ID NO: 2250) AAT-633  Target: 5′-ATCCATGAAGGCTTCCAGGAACTCCTC-3′ (SEQ ID NO: 2390) 5′-GGACACCGAAGAGGCCAAGAAACAG-3′ (SEQ ID NO: 2531) 3′-CCCCUGUGGCUUCUCCGGUUCUUUGUC-5′ (SEQ ID NO: 2251) AAT-801  Target: 5′-GGGGACACCGAAGAGGCCAAGAAACAG-3′ (SEQ ID NO: 2391) 5′-GACACCGAAGAGGCCAAGAAACAGA-3′ (SEQ ID NO: 2532) 3′-CCCUGUGGCUUCUCCGGUUCUUUGUCU-5′ (SEQ ID NO: 2252) AAT-802  Target: 5′-GGGACACCGAAGAGGCCAAGAAACAGA-3′ (SEQ ID NO: 2392) 5′-ACACCGAAGAGGCCAAGAAACAGAU-3′ (SEQ ID NO: 2533) 3′-CCUGUGGCUUCUCCGGUUCUUUGUCUA-5′ (SEQ ID NO: 2253) AAT-803  Target: 5′-GGACACCGAAGAGGCCAAGAAACAGAT-3′ (SEQ ID NO: 2393) 5′-CACCGAAGAGGCCAAGAAACAGAUC-3′ (SEQ ID NO: 2534) 3′-CUGUGGCUUCUCCGGUUCUUUGUCUAG-5′ (SEQ ID NO: 2254) AAT-804  Target: 5′-GACACCGAAGAGGCCAAGAAACAGATC-3′ (SEQ ID NO: 2394) 5′-ACCGAAGAGGCCAAGAAACAGAUCA-3′ (SEQ ID NO: 2535) 3′-UGUGGCUUCUCCGGUUCUUUGUCUAGU-5′ (SEQ ID NO: 2255) AAT-805  Target: 5′-ACACCGAAGAGGCCAAGAAACAGATCA-3′ (SEQ ID NO: 2395) 5′-CCGAAGAGGCCAAGAAACAGAUCAA-3′ (SEQ ID NO: 2536) 3′-GUGGCUUCUCCGGUUCUUUGUCUAGUU-5′ (SEQ ID NO: 2256) AAT-806  Target: 5′-CACCGAAGAGGCCAAGAAACAGATCAA-3′ (SEQ ID NO: 2396) 5′-CGAAGAGGCCAAGAAACAGAUCAAC-3′ (SEQ ID NO: 2537) 3′-UGGCUUCUCCGGUUCUUUGUCUAGUUG-5′ (SEQ ID NO: 2257) AAT-807  Target: 5′-ACCGAAGAGGCCAAGAAACAGATCAAC-3′ (SEQ ID NO: 2397) 5′-GAAGAGGCCAAGAAACAGAUCAACG-3′ (SEQ ID NO: 2538) 3′-GGCUUCUCCGGUUCUUUGUCUAGUUGC-5′ (SEQ ID NO: 2258) AAT-808  Target: 5′-CCGAAGAGGCCAAGAAACAGATCAACG-3′ (SEQ ID NO: 2398) 5′-AAGAGGCCAAGAAACAGAUCAACGA-3′ (SEQ ID NO: 2539) 3′-GCUUCUCCGGUUCUUUGUCUAGUUGCU-5′ (SEQ ID NO: 2259) AAT-809  Target: 5′-CGAAGAGGCCAAGAAACAGATCAACGA-3′ (SEQ ID NO: 2399) 5′-AGAGGCCAAGAAACAGAUCAACGAU-3′ (SEQ ID NO: 2540) 3′-CUUCUCCGGUUCUUUGUCUAGUUGCUA-5′ (SEQ ID NO: 2260) AAT-810  Target: 5′-GAAGAGGCCAAGAAACAGATCAACGAT-3′ (SEQ ID NO: 2400) 5′-GAGGCCAAGAAACAGAUCAACGAUU-3′ (SEQ ID NO: 2541) 3′-UUCUCCGGUUCUUUGUCUAGUUGCUAA-5′ (SEQ ID NO: 2261) AAT-811  Target: 5′-AAGAGGCCAAGAAACAGATCAACGATT-3′ (SEQ ID NO: 2401) 5′-AGGCCAAGAAACAGAUCAACGAUUA-3′ (SEQ ID NO: 2542) 3′-UCUCCGGUUCUUUGUCUAGUUGCUAAU-5′ (SEQ ID NO: 2262) AAT-812  Target: 5′-AGAGGCCAAGAAACAGATCAACGATTA-3′ (SEQ ID NO: 2402) 5′-GGCCAAGAAACAGAUCAACGAUUAC-3′ (SEQ ID NO: 2543) 3′-CUCCGGUUCUUUGUCUAGUUGCUAAUG-5′ (SEQ ID NO: 2263) AAT-813  Target: 5′-GAGGCCAAGAAACAGATCAACGATTAC-3′ (SEQ ID NO: 2403) 5′-UUUUGCUCUGGUGAAUUACAUCUUC-3′ (SEQ ID NO: 2544) 3′-CAAAAACGAGACCACUUAAUGUAGAAG-5′ (SEQ ID NO: 2264) AAT-900  Target: 5′-GTTTTTGCTCTGGTGAATTACATCTTC-3′ (SEQ ID NO: 2404) 5′-UUUGCUCUGGUGAAUUACAUCUUCU-3′ (SEQ ID NO: 2545) 3′-AAAAACGAGACCACUUAAUGUAGAAGA-5′ (SEQ ID NO: 2265) AAT-901  Target: 5′-TTTTTGCTCTGGTGAATTACATCTTCT-3′ (SEQ ID NO: 2405) 5′-UUGCUCUGGUGAAUUACAUCUUCUU-3′ (SEQ ID NO: 2546) 3′-AAAACGAGACCACUUAAUGUAGAAGAA-5′ (SEQ ID NO: 2266) AAT-902  Target: 5′-TTTTGCTCTGGTGAATTACATCTTCTT-3′ (SEQ ID NO: 2406) 5′-UGCUCUGGUGAAUUACAUCUUCUUU-3′ (SEQ ID NO: 2547) 3′-AAACGAGACCACUUAAUGUAGAAGAAA-5′ (SEQ ID NO: 2267) AAT-903  Target: 5′-TTTGCTCTGGTGAATTACATCTTCTTT-3′ (SEQ ID NO: 2407) 5′-GCUCUGGUGAAUUACAUCUUCUUUA-3T (SEQ ID NO: 2548) 3′-AACGAGACCACUUAAUGUAGAAGAAAU-5′ (SEQ ID NO: 2268) AAT-904  Target: 5′-TTGCTCTGGTGAATTACATCTTCTTTA-3′ (SEQ ID NO: 2408) 5′-CUCUGGUGAAUUACAUCUUCUUUAA-3T (SEQ ID NO: 2549) 3′-ACGAGACCACUUAAUGUAGAAGAAAUU-5′ (SEQ ID NO: 2269) AAT-905  Target: 5′-TGCTCTGGTGAATTACATCTTCTTTAA-3′ (SEQ ID NO: 2409) 5′-UCUGGUGAAUUACAUCUUCUUUAAA-3′ (SEQ ID NO: 2550) 3′-CGAGACCACUUAAUGUAGAAGAAAUUU-5T (SEQ ID NO: 2270) AAT-906  Target: 5′-GCTCTGGTGAATTACATCTTCTTTAAA-3′ (SEQ ID NO: 2410) 5′-CUGGUGAAUUACAUCUUCUUUAAAG-3T (SEQ ID NO: 2551) 3′-GAGACCACUUAAUGUAGAAGAAAUUUC-5′ (SEQ ID NO: 2271) AAT-907  Target: 5′-CTCTGGTGAATTACATCTTCTTTAAAG-3′ (SEQ ID NO: 2411) 5′-UGGUGAAUUACAUCUUCUUUAAAGG-3′ (SEQ ID NO: 2552) 3′-AGACCACUUAAUGUAGAAGAAAUUUCC-5′ (SEQ ID NO: 2272) AAT-908  Target: 5′-TCTGGTGAATTACATCTTCTTTAAAGG-3′ (SEQ ID NO: 2412) 5′-GGUGAAUUACAUCUUCUUUAAAGGC-3′ (SEQ ID NO: 2553) 3′-GACCACUUAAUGUAGAAGAAAUUUCCG-5′ (SEQ ID NO: 2273) AAT-909  Target: 5′-CTGGTGAATTACATCTTCTTTAAAGGC-3′ (SEQ ID NO: 2413) 5′-GUGAAUUACAUCUUCUUUAAAGGCA-3′ (SEQ ID NO: 2554) 3′-ACCACUUAAUGUAGAAGAAAUUUCCGU-5′ (SEQ ID NO: 2274) AAT-910  Target: 5′-TGGTGAATTACATCTTCTTTAAAGGCA-3′ (SEQ ID NO: 2414) 5′-UGAAUUACAUCUUCUUUAAAGGCAA-3′ (SEQ ID NO: 2555) 3′-CCACUUAAUGUAGAAGAAAUUUCCGUU-5′ (SEQ ID NO: 2275) AAT-911  Target: 5′-GGTGAATTACATCTTCTTTAAAGGCAA-3′ (SEQ ID NO: 2415) 5′-GAAUUACAUCUUCUUUAAAGGCAAA-3′ (SEQ ID NO: 2556) 3′-CACUUAAUGUAGAAGAAAUUUCCGUUU-5′ (SEQ ID NO: 2276) AAT-912  Target: 5′-GTGAATTACATCTTCTTTAAAGGCAAA-3′ (SEQ ID NO: 2416) 5′-AAUUACAUCUUCUUUAAAGGCAAAU-3′ (SEQ ID NO: 2557) 3′-ACUUAAUGUAGAAGAAAUUUCCGUUUA-5′ (SEQ ID NO: 2277) AAT-913  Target: 5′-TGAATTACATCTTCTTTAAAGGCAAAT-3′ (SEQ ID NO: 2417) 5′-AUUACAUCUUCUUUAAAGGCAAAUG-3′ (SEQ ID NO: 2558) 3′-CUUAAUGUAGAAGAAAUUUCCGUUUAC-5′ (SEQ ID NO: 2278) AAT-914  Target: 5′-GAATTACATCTTCTTTAAAGGCAAATG-3′ (SEQ ID NO: 2418) 5′-UUACAUCUUCUUUAAAGGCAAAUGG-3′ (SEQ ID NO: 2559) 3′-UUAAUGUAGAAGAAAUUUCCGUUUACC-5′ (SEQ ID NO: 2279) AAT-915  Target: 5′-AATTACATCTTCTTTAAAGGCAAATGG-3′ (SEQ ID NO: 2419) 5′-UACAUCUUCUUUAAAGGCAAAUGGG-3′ (SEQ ID NO: 2560) 3′-UAAUGUAGAAGAAAUUUCCGUUUACCC-5′ (SEQ ID NO: 2280) AAT-916  Target: 5′-ATTACATCTTCTTTAAAGGCAAATGGG-3′ (SEQ ID NO: 2420) 5′-ACAUCUUCUUUAAAGGCAAAUGGGA-3′ (SEQ ID NO: 2561) 3′-AAUGUAGAAGAAAUUUCCGUUUACCCU-5′ (SEQ ID NO: 2281) AAT-917  Target: 5′-TTACATCTTCTTTAAAGGCAAATGGGA-3′ (SEQ ID NO: 2421) 5′-CAUCUUCUUUAAAGGCAAAUGGGAG-3′ (SEQ ID NO: 2562) 3′-AUGUAGAAGAAAUUUCCGUUUACCCUC-5′ (SEQ ID NO: 2282) AAT-918  Target: 5′-TACATCTTCTTTAAAGGCAAATGGGAG-3′ (SEQ ID NO: 2422) 5′-UUCUUUAAAGGCAAAUGGGAGAGAC-3′ (SEQ ID NO: 2563) 3′-AGAAGAAAUUUCCGUUUACCCUCUCUG-5′ (SEQ ID NO: 2283) AAT-922  Target: 5′-TCTTCTTTAAAGGCAAATGGGAGAGAC-3′ (SEQ ID NO: 2423) 5′-CUUUAAAGGCAAAUGGGAGAGACCC-3′ (SEQ ID NO: 2564) 3′-AAGAAAUUUCCGUUUACCCUCUCUGGG-5′ (SEQ ID NO: 2284) AAT-924  Target: 5′-TTCTTTAAAGGCAAATGGGAGAGACCC-3′ (SEQ ID NO: 2424) 5′-GCAAAUGGGAGAGACCCUUUGAAGU-3′ (SEQ ID NO: 2565) 3′-UCCGUUUACCCUCUCUGGGAAACUUCA-5′ (SEQ ID NO: 2285) AAT-932  Target: 5′-AGGCAAATGGGAGAGACCCTTTGAAGT-3′ (SEQ ID NO: 2425) 5′-CAAAUGGGAGAGACCCUUUGAAGUC-3′ (SEQ ID NO: 2566) 3′-CCGUUUACCCUCUCUGGGAAACUUCAG-5′ (SEQ ID NO: 2286) AAT-933  Target: 5′-GGCAAATGGGAGAGACCCTTTGAAGTC-3′ (SEQ ID NO: 2426) 5′-AAAUGGGAGAGACCCUUUGAAGUCA-3′ (SEQ ID NO: 2567) 3′-CGUUUACCCUCUCUGGGAAACUUCAGU-5′ (SEQ ID NO: 2287) AAT-934  Target: 5′-GCAAATGGGAGAGACCCTTTGAAGTCA-3′ (SEQ ID NO: 2427) 5′-AAUGGGAGAGACCCUUUGAAGUCAA-3′ (SEQ ID NO: 2568) 3′-GUUUACCCUCUCUGGGAAACUUCAGUU-5′ (SEQ ID NO: 2288) AAT-935  Target: 5′-CAAATGGGAGAGACCCTTTGAAGTCAA-3′ (SEQ ID NO: 2428) 5′-UGUCCAGCUGGGUGCUGCUGAUGAA-3′ (SEQ ID NO: 2569) 3′-CGACAGGUCGACCCACGACGACUACUU-5′ (SEQ ID NO: 2289) AAT-1061 Target: 5′-GCTGTCCAGCTGGGTGCTGCTGATGAA-3′ (SEQ ID NO: 2429) 5′-GUCCAGCUGGGUGCUGCUGAUGAAA-3′ (SEQ ID NO: 2570) 3′-GACAGGUCGACCCACGACGACUACUUU-5′ (SEQ ID NO: 2290) AAT-1062 Target: 5′-CTGTCCAGCTGGGTGCTGCTGATGAAA-3′ (SEQ ID NO: 2430) 5′-UCCAGCUGGGUGCUGCUGAUGAAAU-3′ (SEQ ID NO: 2571) 3′-ACAGGUCGACCCACGACGACUACUUUA-5′ (SEQ ID NO: 2291) AAT-1063 Target: 5′-TGTCCAGCTGGGTGCTGCTGATGAAAT-3′ (SEQ ID NO: 2431) 5′-CCAGCUGGGUGCUGCUGAUGAAAUA-3′ (SEQ ID NO: 2572) 3′-CAGGUCGACCCACGACGACUACUUUAU-5′ (SEQ ID NO: 2292) AAT-1064 Target: 5′-GTCCAGCTGGGTGCTGCTGATGAAATA-3′ (SEQ ID NO: 2432) 5′-CAGCUGGGUGCUGCUGAUGAAAUAC-3′ (SEQ ID NO: 2573) 3′-AGGUCGACCCACGACGACUACUUUAUG-5′ (SEQ ID NO: 2293) AAT-1065 Target: 5′-TCCAGCTGGGTGCTGCTGATGAAATAC-3′ (SEQ ID NO: 2433) 5′-AGCUGGGUGCUGCUGAUGAAAUACC-3′ (SEQ ID NO: 2574) 3′-GGUCGACCCACGACGACUACUUUAUGG-5′ (SEQ ID NO: 2294) AAT-1066 Target: 5′-CCAGCTGGGTGCTGCTGATGAAATACC-3′ (SEQ ID NO: 2434) 5′-GCUGGGUGCUGCUGAUGAAAUACCU-3′ (SEQ ID NO: 2575) 3′-GUCGACCCACGACGACUACUUUAUGGA-5′ (SEQ ID NO: 2295) AAT-1067 Target: 5′-CAGCTGGGTGCTGCTGATGAAATACCT-3′ (SEQ ID NO: 2435) 5′-CUGGGUGCUGCUGAUGAAAUACCUG-3′ (SEQ ID NO: 2576) 3′-UCGACCCACGACGACUACUUUAUGGAC-5′ (SEQ ID NO: 2296) AAT-1068 Target: 5′-AGCTGGGTGCTGCTGATGAAATACCTG-3′ (SEQ ID NO: 2436) 5′-UGGGUGCUGCUGAUGAAAUACCUGG-3′ (SEQ ID NO: 2577) 3′-CGACCCACGACGACUACUUUAUGGACC-5′ (SEQ ID NO: 2297) AAT-1069 Target: 5′-GCTGGGTGCTGCTGATGAAATACCTGG-3′ (SEQ ID NO: 2437) 5′-GGGUGCUGCUGAUGAAAUACCUGGG-3′ (SEQ ID NO: 2578) 3′-GACCCACGACGACUACUUUAUGGACCC-5′ (SEQ ID NO: 2298) AAT-1070 Target: 5′-CTGGGTGCTGCTGATGAAATACCTGGG-3′ (SEQ ID NO: 2438) 5′-GUGCUGCUGAUGAAAUACCUGGGCA-3′ (SEQ ID NO: 2579) 3′-CCCACGACGACUACUUUAUGGACCCGU-5′ (SEQ ID NO: 2299) AAT-1072 Target: 5′-GGGTGCTGCTGATGAAATACCTGGGCA-3′ (SEQ ID NO: 2439) 5′-UGCUGCUGAUGAAAUACCUGGGCAA-3′ (SEQ ID NO: 2580) 3′-CCACGACGACUACUUUAUGGACCCGUU-5′ (SEQ ID NO: 2300) AAT-1073 Target: 5′-GGTGCTGCTGATGAAATACCTGGGCAA-3′ (SEQ ID NO: 2440) 5′-GCUGCUGAUGAAAUACCUGGGCAAU-3′ (SEQ ID NO: 2581) 3′-CACGACGACUACUUUAUGGACCCGUUA-5′ (SEQ ID NO: 2301) AAT-1074 Target: 5′-GTGCTGCTGATGAAATACCTGGGCAAT-3′ (SEQ ID NO: 2441) 5′-CUGCUGAUGAAAUACCUGGGCAAUG-3′ (SEQ ID NO: 2582) 3′-ACGACGACUACUUUAUGGACCCGUUAC-5′ (SEQ ID NO: 2302) AAT-1075 Target: 5′-TGCTGCTGATGAAATACCTGGGCAATG-3′ (SEQ ID NO: 2442) 5′-UGCUGAUGAAAUACCUGGGCAAUGC-3′ (SEQ ID NO: 2583) 3′-CGACGACUACUUUAUGGACCCGUUACG-5′ (SEQ ID NO: 2303) AAT-1076 Target: 5′-GCTGCTGATGAAATACCTGGGCAATGC-3′ (SEQ ID NO: 2443) 5′-GCUGAUGAAAUACCUGGGCAAUGCC-3′ (SEQ ID NO: 2584) 3′-GACGACUACUUUAUGGACCCGUUACGG-5′ (SEQ ID NO: 2304) AAT-1077 Target: 5′-CTGCTGATGAAATACCTGGGCAATGCC-3′ (SEQ ID NO: 2444) 5′-CUGAUGAAAUACCUGGGCAAUGCCA-3′ (SEQ ID NO: 2585) 3′-ACGACUACUUUAUGGACCCGUUACGGU-5′ (SEQ ID NO: 2305) AAT-1078 Target: 5′-TGCTGATGAAATACCTGGGCAATGCCA-3′ (SEQ ID NO: 2445) 5′-UGAUGAAAUACCUGGGCAAUGCCAC-3′ (SEQ ID NO: 2586) 3′-CGACUACUUUAUGGACCCGUUACGGUG-5′ (SEQ ID NO: 2306) AAT-1079 Target: 5′-GCTGATGAAATACCTGGGCAATGCCAC-3′ (SEQ ID NO: 2446) 5′-GAUGAAAUACCUGGGCAAUGCCACC-3′ (SEQ ID NO: 2587) 3′-GACUACUUUAUGGACCCGUUACGGUGG-5′ (SEQ ID NO: 2307) AAT-1080 Target: 5′-CTGATGAAATACCTGGGCAATGCCACC-3′ (SEQ ID NO: 2447) 5′-AUGAAAUACCUGGGCAAUGCCACCG-3′ (SEQ ID NO: 2588) 3′-ACUACUUUAUGGACCCGUUACGGUGGC-5′ (SEQ ID NO: 2308) AAT-1081 Target: 5′-TGATGAAATACCTGGGCAATGCCACCG-3′ (SEQ ID NO: 2448) 5′-GAAAUACCUGGGCAAUGCCACCGCC-3′ (SEQ ID NO: 2589) 3′-UACUUUAUGGACCCGUUACGGUGGCGG-5′ (SEQ ID NO: 2309) AAT-1083 Target: 5′-ATGAAATACCTGGGCAATGCCACCGCC-3′ (SEQ ID NO: 2449) 5′-CAGCACCUGGAAAAUGAACUCACCC-3′ (SEQ ID NO: 2590) 3′-AUGUCGUGGACCUUUUACUUGAGUGGG-5′ (SEQ ID NO: 2310) AAT-1138 Target: 5′-TACAGCACCTGGAAAATGAACTCACCC-3′ (SEQ ID NO: 2450) 5′-CUGGAAAAUGAACUCACCCACGAUA-3′ (SEQ ID NO: 2591) 3′-UGGACCUUUUACUUGAGUGGGUGCUAU-5′ (SEQ ID NO: 2311) AAT-1144 Target: 5′-ACCTGGAAAATGAACTCACCCACGATA-3′ (SEQ ID NO: 2451) 5′-UGGAAAAUGAACUCACCCACGAUAU-3′ (SEQ ID NO: 2592) 3′-GGACCUUUUACUUGAGUGGGUGCUAUA-5′ (SEQ ID NO: 2312) AAT-1145 Target: 5′-CCTGGAAAATGAACTCACCCACGATAT-3′ (SEQ ID NO: 2452) 5′-GAUAUCAUCACCAAGUUCCUGGAAA-3′ (SEQ ID NO: 2593) 3′-UGCUAUAGUAGUGGUUCAAGGACCUUU-5′ (SEQ ID NO: 2313) AAT-1165 Target: 5′-ACGATATCATCACCAAGTTCCTGGAAA-3′ (SEQ ID NO: 2453) 5′-CAAGUUCCUGGAAAAUGAAGACAGA-3′ (SEQ ID NO: 2594) 3′-UGGUUCAAGGACCUUUUACUUCUGUCU-5′ (SEQ ID NO: 2314) AAT-1176 Target: 5′-ACCAAGTTCCTGGAAAATGAAGACAGA-3′ (SEQ ID NO: 2454) 5′-CCAUUACUGGAACCUAUGAUCUGAA-3′ (SEQ ID NO: 2595) 3′-CAGGUAAUGACCUUGGAUACUAGACUU-5′ (SEQ ID NO: 2315) AAT-1232 Target: 5′-GTCCATTACTGGAACCTATGATCTGAA-3′ (SEQ ID NO: 2455) 5′-CAUUACUGGAACCUAUGAUCUGAAG-3′ (SEQ ID NO: 2596) 3′-AGGUAAUGACCUUGGAUACUAGACUUC-5′ (SEQ ID NO: 2316) AAT-1233 Target: 5′-TCCATTACTGGAACCTATGATCTGAAG-3′ (SEQ ID NO: 2456) 5′-AUUACUGGAACCUAUGAUCUGAAGA-3′ (SEQ ID NO: 2597) 3′-GGUAAUGACCUUGGAUACUAGACUUCU-5′ (SEQ ID NO: 2317) AAT-1234 Target: 5′-CCATTACTGGAACCTATGATCTGAAGA-3′ (SEQ ID NO: 2457) 5′-UUACUGGAACCUAUGAUCUGAAGAG-3′ (SEQ ID NO: 2598) 3′-GUAAUGACCUUGGAUACUAGACUUCUC-5′ (SEQ ID NO: 2318) AAT-1235 Target: 5′-CATTACTGGAACCTATGATCTGAAGAG-3′ (SEQ ID NO: 2458) 5′-UACUGGAACCUAUGAUCUGAAGAGC-3′ (SEQ ID NO: 2599) 3′-UAAUGACCUUGGAUACUAGACUUCUCG-5′ (SEQ ID NO: 2319) AAT-1236 Target: 5′-ATTACTGGAACCTATGATCTGAAGAGC-3′ (SEQ ID NO: 2459) 5′-ACUGGAACCUAUGAUCUGAAGAGCG-3′ (SEQ ID NO: 2600) 3′-AAUGACCUUGGAUACUAGACUUCUCGC-5′ (SEQ ID NO: 2320) AAT-1237 Target: 5′-TTACTGGAACCTATGATCTGAAGAGCG-3′ (SEQ ID NO: 2460) 5′-CUGGAACCUAUGAUCUGAAGAGCGU-3′ (SEQ ID NO: 2601) 3′-AUGACCUUGGAUACUAGACUUCUCGCA-5′ (SEQ ID NO: 2321) AAT-1238 Target: 5′-TACTGGAACCTATGATCTGAAGAGCGT-3′ (SEQ ID NO: 2461) 5′-UGGAACCUAUGAUCUGAAGAGCGUC-3′ (SEQ ID NO: 2602) 3′-UGACCUUGGAUACUAGACUUCUCGCAG-5′ (SEQ ID NO: 2322) AAT-1239 Target: 5′-ACTGGAACCTATGATCTGAAGAGCGTC-3′ (SEQ ID NO: 2462) 5′-GGAACCUAUGAUCUGAAGAGCGUCC-3′ (SEQ ID NO: 2603) 3′-GACCUUGGAUACUAGACUUCUCGCAGG-5′ (SEQ ID NO: 2323) AAT-1240 Target: 5′-CTGGAACCTATGATCTGAAGAGCGTCC-3′ (SEQ ID NO: 2463) 5′-AUCACUAAGGUCUUCAGCAAUGGGG-3′ (SEQ ID NO: 2604) 3′-CGUAGUGAUUCCAGAAGUCGUUACCCC-5′ (SEQ ID NO: 2324) AAT-1279 Target: 5′-GCATCACTAAGGTCTTCAGCAATGGGG-3′ (SEQ ID NO: 2464) 5′-UCACUAAGGUCUUCAGCAAUGGGGC-3′ (SEQ ID NO: 2605) 3′-GUAGUGAUUCCAGAAGUCGUUACCCCG-5′ (SEQ ID NO: 2325) AAT-1280 Target: 5′-CATCACTAAGGTCTTCAGCAATGGGGC-3′ (SEQ ID NO: 2465) 5′-CACUAAGGUCUUCAGCAAUGGGGCU-3′ (SEQ ID NO: 2606) 3′-UAGUGAUUCCAGAAGUCGUUACCCCGA-5′ (SEQ ID NO: 2326) AAT-1281 Target: 5′-ATCACTAAGGTCTTCAGCAATGGGGCT-3′ (SEQ ID NO: 2466) 5′-CUAAGGUCUUCAGCAAUGGGGCUGA-3′ (SEQ ID NO: 2607) 3′-GUGAUUCCAGAAGUCGUUACCCCGACU-5′ (SEQ ID NO: 2327) AAT-1283 Target: 5′-CACTAAGGTCTTCAGCAATGGGGCTGA-3′ (SEQ ID NO: 2467) 5′-UAAGGUCUUCAGCAAUGGGGCUGAC-3′ (SEQ ID NO: 2608) 3′-UGAUUCCAGAAGUCGUUACCCCGACUG-5′ (SEQ ID NO: 2328) AAT-1284 Target: 5′-ACTAAGGTCTTCAGCAATGGGGCTGAC-3′ (SEQ ID NO: 2468) 5′-UGAAGCUCUCCAAGGCCGUGCAUAA-3′ (SEQ ID NO: 2609) 3′-GGACUUCGAGAGGUUCCGGCACGUAUU-5′ (SEQ ID NO: 2329) AAT-1337 Target: 5′-CCTGAAGCTCTCCAAGGCCGTGCATAA-3′ (SEQ ID NO: 2469) 5′-GAAGCUCUCCAAGGCCGUGCAUAAG-3′ (SEQ ID NO: 2610) 3′-GACUUCGAGAGGUUCCGGCACGUAUUC-5′ (SEQ ID NO: 2330) AAT-1338 Target: 5′-CTGAAGCTCTCCAAGGCCGTGCATAAG-3′ (SEQ ID NO: 2470) 5′-AAGCUCUCCAAGGCCGUGCAUAAGG-3′ (SEQ ID NO: 2611) 3′-ACUUCGAGAGGUUCCGGCACGUAUUCC-5′ (SEQ ID NO: 2331) AAT-1339 Target: 5′-TGAAGCTCTCCAAGGCCGTGCATAAGG-3′ (SEQ ID NO: 2471) 5′-CCGAGGUCAAGUUCAACAAACCCUU-3′ (SEQ ID NO: 2612) 3′-GGGGCUCCAGUUCAAGUUGUUUGGGAA-5′ (SEQ ID NO: 2332) AAT-1442 Target: 5′-CCCCGAGGTCAAGTTCAACAAACCCTT-3′ (SEQ ID NO: 2472) 5′-CGAGGUCAAGUUCAACAAACCCUUU-3′ (SEQ ID NO: 2613) 3′-GGGCUCCAGUUCAAGUUGUUUGGGAAA-5′ (SEQ ID NO: 2333) AAT-1443 Target: 5′-CCCGAGGTCAAGTTCAACAAACCCTTT-3′ (SEQ ID NO: 2473) 5′-GAGGUCAAGUUCAACAAACCCUUUG-3′ (SEQ ID NO: 2614) 3′-GGCUCCAGUUCAAGUUGUUUGGGAAAC-5′ (SEQ ID NO: 2334) AAT-1444 Target: 5′-CCGAGGTCAAGTTCAACAAACCCTTTG-3′ (SEQ ID NO: 2474) 5′-AGGUCAAGUUCAACAAACCCUUUGU-3′ (SEQ ID NO: 2615) 3′-GCUCCAGUUCAAGUUGUUUGGGAAACA-5′ (SEQ ID NO: 2335) AAT-1445 Target: 5′-CGAGGTCAAGTTCAACAAACCCTTTGT-3′ (SEQ ID NO: 2475) 5′-GGUCAAGUUCAACAAACCCUUUGUC-3′ (SEQ ID NO: 2616) 3′-CUCCAGUUCAAGUUGUUUGGGAAACAG-5′ (SEQ ID NO: 2336) AAT-1446 Target: 5′-GAGGTCAAGTTCAACAAACCCTTTGTC-3′ (SEQ ID NO: 2476) 5′-GUCAAGUUCAACAAACCCUUUGUCU-3′ (SEQ ID NO: 2617) 3′-UCCAGUUCAAGUUGUUUGGGAAACAGA-5′ (SEQ ID NO: 2337) AAT-1447 Target: 5′-AGGTCAAGTTCAACAAACCCTTTGTCT-3′ (SEQ ID NO: 2477) 5′-UCAAGUUCAACAAACCCUUUGUCUU-3′ (SEQ ID NO: 2618) 3′-CCAGUUCAAGUUGUUUGGGAAACAGAA-5′ (SEQ ID NO: 2338) AAT-1448 Target: 5′-GGTCAAGTTCAACAAACCCTTTGTCTT-3′ (SEQ ID NO: 2478) 5′-CAAGUUCAACAAACCCUUUGUCUUC-3′ (SEQ ID NO: 2619) 3′-CAGUUCAAGUUGUUUGGGAAACAGAAG-5′ (SEQ ID NO: 2339) AAT-1449 Target: 5′-GTCAAGTTCAACAAACCCTTTGTCTTC-3′ (SEQ ID NO: 2479) 5′-AAGUUCAACAAACCCUUUGUCUUCU-3′ (SEQ ID NO: 2620) 3′-AGUUCAAGUUGUUUGGGAAACAGAAGA-5′ (SEQ ID NO: 2340) AAT-1450 Target: 5′-TCAAGTTCAACAAACCCTTTGTCTTCT-3′ (SEQ ID NO: 2480) 5′-AGUUCAACAAACCCUUUGUCUUCUU-3′ (SEQ ID NO: 2621) 3′-GUUCAAGUUGUUUGGGAAACAGAAGAA-5′ (SEQ ID NO: 2341) AAT-1451 Target: 5′-CAAGTTCAACAAACCCTTTGTCTTCTT-3′ (SEQ ID NO: 2481) 

TABLE 14  Further DsiRNA Target Sequences (21mers) in a-1 antitrypsin mRNA AAT-366 21 nt Target: 5′-CCCCAGGGAGAUGCUGCCCAG-3′ (SEQ ID NO: 2622) AAT-367 21 nt Target: 5′-CCCAGGGAGAUGCUGCCCAGA-3′ (SEQ ID NO: 2623) AAT-368 21 nt Target: 5′-CCAGGGAGAUGCUGCCCAGAA-3′ (SEQ ID NO: 2624) AAT-369 21 nt Target: 5′-CAGGGAGAUGCUGCCCAGAAG-3′ (SEQ ID NO: 2625) AAT-370 21 nt Target: 5′-AGGGAGAUGCUGCCCAGAAGA-3′ (SEQ ID NO: 2626) AAT-371 21 nt Target: 5′-GGGAGAUGCUGCCCAGAAGAC-3′ (SEQ ID NO: 2627) AAT-391 21 nt Target: 5′-CAGAUACAUCCCACCAUGAUC-3′ (SEQ ID NO: 2628) AAT-392 21 nt Target: 5′-AGAUACAUCCCACCAUGAUCA-3′ (SEQ ID NO: 2629) AAT-393 21 nt Target: 5′-GAUACAUCCCACCAUGAUCAG-3′ (SEQ ID NO: 2630) AAT-394 21 nt Target: 5′-AUACAUCCCACCAUGAUCAGG-3′ (SEQ ID NO: 2631) AAT-485 21 nt Target: 5′-ACACCAGUCCAACAGCACCAA-3′ (SEQ ID NO: 2632) AAT-486 21 nt Target: 5′-CACCAGUCCAACAGCACCAAU-3′ (SEQ ID NO: 2633) AAT-487 21 nt Target: 5′-ACCAGUCCAACAGCACCAAUA-3′ (SEQ ID NO: 2634) AAT-488 21 nt Target: 5′-CCAGUCCAACAGCACCAAUAU-3′ (SEQ ID NO: 2635) AAT-489 21 nt Target: 5′-CAGUCCAACAGCACCAAUAUC-3′ (SEQ ID NO: 2636) AAT-490 21 nt Target: 5′-AGUCCAACAGCACCAAUAUCU-3′ (SEQ ID NO: 2637) AAT-491 21 nt Target: 5′-GUCCAACAGCACCAAUAUCUU-3′ (SEQ ID NO: 2638) AAT-492 21 nt Target: 5′-UCCAACAGCACCAAUAUCUUC-3′ (SEQ ID NO: 2639) AAT-493 21 nt Target: 5′-CCAACAGCACCAAUAUCUUCU-3′ (SEQ ID NO: 2640) AAT-494 21 nt Target: 5′-CAACAGCACCAAUAUCUUCUU-3′ (SEQ ID NO: 2641) AAT-495 21 nt Target: 5′-AACAGCACCAAUAUCUUCUUC-3′ (SEQ ID NO: 2642) AAT-496 21 nt Target: 5′-ACAGCACCAAUAUCUUCUUCU-3′ (SEQ ID NO: 2643) AAT-497 21 nt Target: 5′-CAGCACCAAUAUCUUCUUCUC-3′ (SEQ ID NO: 2644) AAT-498 21 nt Target: 5′-AGCACCAAUAUCUUCUUCUCC-3′ (SEQ ID NO: 2645) AAT-499 21 nt Target: 5′-GCACCAAUAUCUUCUUCUCCC-3′ (SEQ ID NO: 2646) AAT-516 21 nt Target: 5′-UCCCCAGUGAGCAUCGCUACA-3′ (SEQ ID NO: 2647) AAT-517 21 nt Target: 5′-CCCCAGUGAGCAUCGCUACAG-3′ (SEQ ID NO: 2648) AAT-518 21 nt Target: 5′-CCCAGUGAGCAUCGCUACAGC-3′ (SEQ ID NO: 2649) AAT-519 21 nt Target: 5′-CCAGUGAGCAUCGCUACAGCC-3′ (SEQ ID NO: 2650) AAT-520 21 nt Target: 5′-CAGUGAGCAUCGCUACAGCCU-3′ (SEQ ID NO: 2651) AAT-521 21 nt Target: 5′-AGUGAGCAUCGCUACAGCCUU-3′ (SEQ ID NO: 2652) AAT-522 21 nt Target: 5′-GUGAGCAUCGCUACAGCCUUU-3′ (SEQ ID NO: 2653) AAT-523 21 nt Target: 5′-UGAGCAUCGCUACAGCCUUUG-3′ (SEQ ID NO: 2654) AAT-524 21 nt Target: 5′-GAGCAUCGCUACAGCCUUUGC-3′ (SEQ ID NO: 2655) AAT-525 21 nt Target: 5′-AGCAUCGCUACAGCCUUUGCA-3′ (SEQ ID NO: 2656) AAT-526 21 nt Target: 5′-GCAUCGCUACAGCCUUUGCAA-3′ (SEQ ID NO: 2657) AAT-527 21 nt Target: 5′-CAUCGCUACAGCCUUUGCAAU-3′ (SEQ ID NO: 2658) AAT-528 21 nt Target: 5′-AUCGCUACAGCCUUUGCAAUG-3′ (SEQ ID NO: 2659) AAT-529 21 nt Target: 5′-UCGCUACAGCCUUUGCAAUGC-3′ (SEQ ID NO: 2660) AAT-530 21 nt Target: 5′-CGCUACAGCCUUUGCAAUGCU-3′ (SEQ ID NO: 2661) AAT-531 21 nt Target: 5′-GCUACAGCCUUUGCAAUGCUC-3′ (SEQ ID NO: 2662) AAT-552 21 nt Target: 5′-UCCCUGGGGACCAAGGCUGAC-3′ (SEQ ID NO: 2663) AAT-556 21 nt Target: 5′-UGGGGACCAAGGCUGACACUC-3′ (SEQ ID NO: 2664) AAT-557 21 nt Target: 5′-GGGGACCAAGGCUGACACUCA-3′ (SEQ ID NO: 2665) AAT-558 21 nt Target: 5′-GGGACCAAGGCUGACACUCAC-3′ (SEQ ID NO: 2666) AAT-579 21 nt Target: 5′-GAUGAAAUCCUGGAGGGCCUG-3′ (SEQ ID NO: 2667) AAT-580 21 nt Target: 5′-AUGAAAUCCUGGAGGGCCUGA-3′ (SEQ ID NO: 2668) AAT-632 21 nt Target: 5′-GAUCCAUGAAGGCUUCCAGGA-3′ (SEQ ID NO: 2669) AAT-633 21 nt Target: 5′-AUCCAUGAAGGCUUCCAGGAA-3′ (SEQ ID NO: 2670) AAT-801 21 nt Target: 5′-GGGGACACCGAAGAGGCCAAG-3′ (SEQ ID NO: 2671) AAT-802 21 nt Target: 5′-GGGACACCGAAGAGGCCAAGA-3′ (SEQ ID NO: 2672) AAT-803 21 nt Target: 5′-GGACACCGAAGAGGCCAAGAA-3′ (SEQ ID NO: 2673) AAT-804 21 nt Target: 5′-GACACCGAAGAGGCCAAGAAA-3′ (SEQ ID NO: 2674) AAT-805 21 nt Target: 5′-ACACCGAAGAGGCCAAGAAAC-3′ (SEQ ID NO: 2675) AAT-806 21 nt Target: 5′-CACCGAAGAGGCCAAGAAACA-3′ (SEQ ID NO: 2676) AAT-807 21 nt Target: 5′-ACCGAAGAGGCCAAGAAACAG-3′ (SEQ ID NO: 2677) AAT-808 21 nt Target: 5′-CCGAAGAGGCCAAGAAACAGA-3′ (SEQ ID NO: 2678) AAT-809 21 nt Target: 5′-CGAAGAGGCCAAGAAACAGAU-3′ (SEQ ID NO: 2679) AAT-810 21 nt Target: 5′-GAAGAGGCCAAGAAACAGAUC-3′ (SEQ ID NO: 2680) AAT-811 21 nt Target: 5′-AAGAGGCCAAGAAACAGAUCA-3′ (SEQ ID NO: 2681) AAT-812 21 nt Target: 5′-AGAGGCCAAGAAACAGAUCAA-3′ (SEQ ID NO: 2682) AAT-813 21 nt Target: 5′-GAGGCCAAGAAACAGAUCAAC-3′ (SEQ ID NO: 2683) AAT-900 21 nt Target: 5′-GUUUUUGCUCUGGUGAAUUAC-3′ (SEQ ID NO: 2684) AAT-901 21 nt Target: 5′-UUUUUGCUCUGGUGAAUUACA-3′ (SEQ ID NO: 2685) AAT-902 21 nt Target: 5′-UUUUGCUCUGGUGAAUUACAU-3′ (SEQ ID NO: 2686) AAT-903 21 nt Target: 5′-UUUGCUCUGGUGAAUUACAUC-3′ (SEQ ID NO: 2687) AAT-904 21 nt Target: 5′-UUGCUCUGGUGAAUUACAUCU-3′ (SEQ ID NO: 2688) AAT-905 21 nt Target: 5′-UGCUCUGGUGAAUUACAUCUU-3′ (SEQ ID NO: 2689) AAT-906 21 nt Target: 5′-GCUCUGGUGAAUUACAUCUUC-3′ (SEQ ID NO: 2690) AAT-907 21 nt Target: 5′-CUCUGGUGAAUUACAUCUUCU-3′ (SEQ ID NO: 2691) AAT-908 21 nt Target: 5′-UCUGGUGAAUUACAUCUUCUU-3′ (SEQ ID NO: 2692) AAT-909 21 nt Target: 5′-CUGGUGAAUUACAUCUUCUUU-3′ (SEQ ID NO: 2693) AAT-910 21 nt Target: 5′-UGGUGAAUUACAUCUUCUUUA-3′ (SEQ ID NO: 2694) AAT-911 21 nt Target: 5′-GGUGAAUUACAUCUUCUUUAA-3′ (SEQ ID NO: 2695) AAT-912 21 nt Target: 5′-GUGAAUUACAUCUUCUUUAAA-3′ (SEQ ID NO: 2696) AAT-913 21 nt Target: 5′-UGAAUUACAUCUUCUUUAAAG-3′ (SEQ ID NO: 2697) AAT-914 21 nt Target: 5′-GAAUUACAUCUUCUUUAAAGG-3′ (SEQ ID NO: 2698) AAT-915 21 nt Target: 5′-AAUUACAUCUUCUUUAAAGGC-3′ (SEQ ID NO: 2699) AAT-916 21 nt Target: 5′-AUUACAUCUUCUUUAAAGGCA-3′ (SEQ ID NO: 2700) AAT-917 21 nt Target: 5′-UUACAUCUUCUUUAAAGGCAA-3′ (SEQ ID NO: 2701) AAT-918 21 nt Target: 5′-UACAUCUUCUUUAAAGGCAAA-3′ (SEQ ID NO: 2702) AAT-922 21 nt Target: 5′-UCUUCUUUAAAGGCAAAUGGG-3′ (SEQ ID NO: 2703) AAT-924 21 nt Target: 5′-UUCUUUAAAGGCAAAUGGGAG-3′ (SEQ ID NO: 2704) AAT-932 21 nt Target: 5′-AGGCAAAUGGGAGAGACCCUU-3′ (SEQ ID NO: 2705) AAT-933 21 nt Target: 5′-GGCAAAUGGGAGAGACCCUUU-3′ (SEQ ID NO: 2706) AAT-934 21 nt Target: 5′-GCAAAUGGGAGAGACCCUUUG-3′ (SEQ ID NO: 2707) AAT-935 21 nt Target: 5′-CAAAUGGGAGAGACCCUUUGA-3′ (SEQ ID NO: 2708) AAT-1061 21 nt Target: 5′-GCUGUCCAGCUGGGUGCUGCU-3′ (SEQ ID NO: 2709) AAT-1062 21 nt Target: 5′-CUGUCCAGCUGGGUGCUGCUG-3′ (SEQ ID NO: 2710) AAT-1063 21 nt Target: 5′-UGUCCAGCUGGGUGCUGCUGA-3′ (SEQ ID NO: 2711) AAT-1064 21 nt Target: 5′-GUCCAGCUGGGUGCUGCUGAU-3′ (SEQ ID NO: 2712) AAT-1065 21 nt Target: 5′-UCCAGCUGGGUGCUGCUGAUG-3′ (SEQ ID NO: 2713) AAT-1066 21 nt Target: 5′-CCAGCUGGGUGCUGCUGAUGA-3′ (SEQ ID NO: 2714) AAT-1067 21 nt Target: 5′-CAGCUGGGUGCUGCUGAUGAA-3′ (SEQ ID NO: 2715) AAT-1068 21 nt Target: 5′-AGCUGGGUGCUGCUGAUGAAA-3′ (SEQ ID NO: 2716) AAT-1069 21 nt Target: 5′-GCUGGGUGCUGCUGAUGAAAU-3′ (SEQ ID NO: 2717) AAT-1070 21 nt Target: 5′-CUGGGUGCUGCUGAUGAAAUA-3′ (SEQ ID NO: 2718) AAT-1072 21 nt Target: 5′-GGGUGCUGCUGAUGAAAUACC-3′ (SEQ ID NO: 2719) AAT-1073 21 nt Target: 5′-GGUGCUGCUGAUGAAAUACCU-3′ (SEQ ID NO: 2720) AAT-1074 21 nt Target: 5′-GUGCUGCUGAUGAAAUACCUG-3′ (SEQ ID NO: 2721) AAT-1075 21 nt Target: 5′-UGCUGCUGAUGAAAUACCUGG-3′ (SEQ ID NO: 2722) AAT-1076 21 nt Target: 5′-GCUGCUGAUGAAAUACCUGGG-3′ (SEQ ID NO: 2723) AAT-1077 21 nt Target: 5′-CUGCUGAUGAAAUACCUGGGC-3′ (SEQ ID NO: 2724) AAT-1078 21 nt Target: 5′-UGCUGAUGAAAUACCUGGGCA-3′ (SEQ ID NO: 2725) AAT-1079 21 nt Target: 5′-GCUGAUGAAAUACCUGGGCAA-3′ (SEQ ID NO: 2726) AAT-1080 21 nt Target: 5′-CUGAUGAAAUACCUGGGCAAU-3′ (SEQ ID NO: 2727) AAT-1081 21 nt Target: 5′-UGAUGAAAUACCUGGGCAAUG-3′ (SEQ ID NO: 2728) AAT-1083 21 nt Target: 5′-AUGAAAUACCUGGGCAAUGCC-3′ (SEQ ID NO: 2729) AAT-1138 21 nt Target: 5′-UACAGCACCUGGAAAAUGAAC-3′ (SEQ ID NO: 2730) AAT-1144 21 nt Target: 5′-ACCUGGAAAAUGAACUCACCC-3′ (SEQ ID NO: 2731) AAT-1145 21 nt Target: 5′-CCUGGAAAAUGAACUCACCCA-3′ (SEQ ID NO: 2732) AAT-1165 21 nt Target: 5′-ACGAUAUCAUCACCAAGUUCC-3′ (SEQ ID NO: 2733) AAT-1176 21 nt Target: 5′-ACCAAGUUCCUGGAAAAUGAA-3′ (SEQ ID NO: 2734) AAT-1232 21 nt Target: 5′-GUCCAUUACUGGAACCUAUGA-3′ (SEQ ID NO: 2735) AAT-1233 21 nt Target: 5′-UCCAUUACUGGAACCUAUGAU-3′ (SEQ ID NO: 2736) AAT-1234 21 nt Target: 5′-CCAUUACUGGAACCUAUGAUC-3′ (SEQ ID NO: 2737) AAT-1235 21 nt Target: 5′-CAUUACUGGAACCUAUGAUCU-3′ (SEQ ID NO: 2738) AAT-1236 21 nt Target: 5′-AUUACUGGAACCUAUGAUCUG-3′ (SEQ ID NO: 2739) AAT-1237 21 nt Target: 5′-UUACUGGAACCUAUGAUCUGA-3′ (SEQ ID NO: 2740) AAT-1238 21 nt Target: 5′-UACUGGAACCUAUGAUCUGAA-3′ (SEQ ID NO: 2741) AAT-1239 21 nt Target: 5′-ACUGGAACCUAUGAUCUGAAG-3′ (SEQ ID NO: 2742) AAT-1240 21 nt Target: 5′-CUGGAACCUAUGAUCUGAAGA-3′ (SEQ ID NO: 2743) AAT-1279 21 nt Target: 5′-GCAUCACUAAGGUCUUCAGCA-3′ (SEQ ID NO: 2744) AAT-1280 21 nt Target: 5′-CAUCACUAAGGUCUUCAGCAA-3′ (SEQ ID NO: 2745) AAT-1281 21 nt Target: 5′-AUCACUAAGGUCUUCAGCAAU-3′ (SEQ ID NO: 2746) AAT-1283 21 nt Target: 5′-CACUAAGGUCUUCAGCAAUGG-3′ (SEQ ID NO: 2747) AAT-1284 21 nt Target: 5′-ACUAAGGUCUUCAGCAAUGGG-3′ (SEQ ID NO: 2748) AAT-1337 21 nt Target: 5′-CCUGAAGCUCUCCAAGGCCGU-3′ (SEQ ID NO: 2749) AAT-1338 21 nt Target: 5′-CUGAAGCUCUCCAAGGCCGUG-3′ (SEQ ID NO: 2750) AAT-1339 21 nt Target: 5′-UGAAGCUCUCCAAGGCCGUGC-3′ (SEQ ID NO: 2751) AAT-1442 21 nt Target: 5′-CCCCGAGGUCAAGUUCAACAA-3′ (SEQ ID NO: 2752) AAT-1443 21 nt Target: 5′-CCCGAGGUCAAGUUCAACAAA-3′ (SEQ ID NO: 2753) AAT-1444 21 nt Target: 5′-CCGAGGUCAAGUUCAACAAAC-3′ (SEQ ID NO: 2754) AAT-1445 21 nt Target: 5′-CGAGGUCAAGUUCAACAAACC-3′ (SEQ ID NO: 2755) AAT-1446 21 nt Target: 5′-GAGGUCAAGUUCAACAAACCC-3′ (SEQ ID NO: 2756) AAT-1447 21 nt Target: 5′-AGGUCAAGUUCAACAAACCCU-3′ (SEQ ID NO: 2757) AAT-1448 21 nt Target: 5′-GGUCAAGUUCAACAAACCCUU-3′ (SEQ ID NO: 2758) AAT-1449 21 nt Target: 5′-GUCAAGUUCAACAAACCCUUU-3′ (SEQ ID NO: 2759) AAT-1450 21 nt Target: 5′-UCAAGUUCAACAAACCCUUUG-3′ (SEQ ID NO: 2760) AAT-1451 21 nt Target: 5′-CAAGUUCAACAAACCCUUUGU-3′ (SEQ ID NO: 2761)

TABLE 15  Further Human Anti-α-1 antitrypsin “Blunt/Blunt” DsiRNAs 5′-CCCCAGGGAGAUGCUGCCCAGAAGACA-3′ (SEQ ID NO: 2762) 3′-GGGGUCCCUCUACGACGGGUCUUCUGU-5′ (SEQ ID NO: 2202) AAT-366  Target: 5′-CCCCAGGGAGATGCTGCCCAGAAGACA-3′ (SEQ ID NO: 2342) 5′-CCCAGGGAGAUGCUGCCCAGAAGACAG-3′ (SEQ ID NO: 2763) 3′-GGGUCCCUCUACGACGGGUCUUCUGUC-5′ (SEQ ID NO: 2203) AAT-367  Target: 5′-CCCAGGGAGATGCTGCCCAGAAGACAG-3′ (SEQ ID NO: 2343) 5′-CCAGGGAGAUGCUGCCCAGAAGACAGA-3′ (SEQ ID NO: 2764) 3′-GGUCCCUCUACGACGGGUCUUCUGUCU-5′ (SEQ ID NO: 2204) AAT-368  Target: 5′-CCAGGGAGATGCTGCCCAGAAGACAGA-3′ (SEQ ID NO: 2344) 5′-CAGGGAGAUGCUGCCCAGAAGACAGAU-3′ (SEQ ID NO: 2765) 3′-GUCCCUCUACGACGGGUCUUCUGUCUA-5′ (SEQ ID NO: 2205) AAT-369  Target: 5′-CAGGGAGATGCTGCCCAGAAGACAGAT-3′ (SEQ ID NO: 2345) 5′-AGGGAGAUGCUGCCCAGAAGACAGAUA-3′ (SEQ ID NO: 2766) 3′-UCCCUCUACGACGGGUCUUCUGUCUAU-5′ (SEQ ID NO: 2206) AAT-370  Target: 5′-AGGGAGATGCTGCCCAGAAGACAGATA-3′ (SEQ ID NO: 2346) 5′-GGGAGAUGCUGCCCAGAAGACAGAUAC-3′ (SEQ ID NO: 2767) 3′-CCCUCUACGACGGGUCUUCUGUCUAUG-5′ (SEQ ID NO: 2207) AAT-371  Target: 5′-GGGAGATGCTGCCCAGAAGACAGATAC-3′ (SEQ ID NO: 2347) 5′-CAGAUACAUCCCACCAUGAUCAGGAUC-3′ (SEQ ID NO: 2768) 3′-GUCUAUGUAGGGUGGUACUAGUCCUAG-5′ (SEQ ID NO: 2208) AAT-391  Target: 5′-CAGATACATCCCACCATGATCAGGATC-3′ (SEQ ID NO: 2348) 5′-AGAUACAUCCCACCAUGAUCAGGAUCA-3′ (SEQ ID NO: 2769) 3′-UCUAUGUAGGGUGGUACUAGUCCUAGU-5′ (SEQ ID NO: 2209) AAT-392  Target: 5′-AGATACATCCCACCATGATCAGGATCA-3′ (SEQ ID NO: 2349) 5′-GAUACAUCCCACCAUGAUCAGGAUCAC-3′ (SEQ ID NO: 2770) 3′-CUAUGUAGGGUGGUACUAGUCCUAGUG-5′ (SEQ ID NO: 2210) AAT-393  Target: 5′-GATACATCCCACCATGATCAGGATCAC-3′ (SEQ ID NO: 2350) 5′-AUACAUCCCACCAUGAUCAGGAUCACC-3′ (SEQ ID NO: 2771) 3′-UAUGUAGGGUGGUACUAGUCCUAGUGG-5′ (SEQ ID NO: 2211) AAT-394  Target: 5′-ATACATCCCACCATGATCAGGATCACC-3′ (SEQ ID NO: 2351) 5′-ACACCAGUCCAACAGCACCAAUAUCUU-3′ (SEQ ID NO: 2772) 3′-UGUGGUCAGGUUGUCGUGGUUAUAGAA-5′ (SEQ ID NO: 2212) AAT-485  Target: 5′-ACACCAGTCCAACAGCACCAATATCTT-3′ (SEQ ID NO: 2352) 5′-CACCAGUCCAACAGCACCAAUAUCUUC-3′ (SEQ ID NO: 2773) 3′-GUGGUCAGGUUGUCGUGGUUAUAGAAG-5′ (SEQ ID NO: 2213) AAT-486  Target: 5′-CACCAGTCCAACAGCACCAATATCTTC-3′ (SEQ ID NO: 2353) 5′-ACCAGUCCAACAGCACCAAUAUCUUCU-3′ (SEQ ID NO: 2774) 3′-UGGUCAGGUUGUCGUGGUUAUAGAAGA-5′ (SEQ ID NO: 2214) AAT-487  Target: 5′-ACCAGTCCAACAGCACCAATATCTTCT-3′ (SEQ ID NO: 2354) 5′-CCAGUCCAACAGCACCAAUAUCUUCUU-3′ (SEQ ID NO: 2775) 3′-GGUCAGGUUGUCGUGGUUAUAGAAGAA-5′ (SEQ ID NO: 2215) AAT-488  Target: 5′-CCAGTCCAACAGCACCAATATCTTCTT-3′ (SEQ ID NO: 2355) 5′-CAGUCCAACAGCACCAAUAUCUUCUUC-3′ (SEQ ID NO: 2776) 3′-GUCAGGUUGUCGUGGUUAUAGAAGAAG-5′ (SEQ ID NO: 2216) AAT-489  Target: 5′-CAGTCCAACAGCACCAATATCTTCTTC-3′ (SEQ ID NO: 2356) 5′-AGUCCAACAGCACCAAUAUCUUCUUCU-3′ (SEQ ID NO: 2777) 3′-UCAGGUUGUCGUGGUUAUAGAAGAAGA-5′ (SEQ ID NO: 2217) AAT-490  Target: 5′-AGTCCAACAGCACCAATATCTTCTTCT-3′ (SEQ ID NO: 2357) 5′-GUCCAACAGCACCAAUAUCUUCUUCUC-3′ (SEQ ID NO: 2778) 3′-CAGGUUGUCGUGGUUAUAGAAGAAGAG-5′ (SEQ ID NO: 2218) AAT-491  Target: 5′-GTCCAACAGCACCAATATCTTCTTCTC-3′ (SEQ ID NO: 2358) 5′-UCCAACAGCACCAAUAUCUUCUUCUCC-3′ (SEQ ID NO: 2779) 3′-AGGUUGUCGUGGUUAUAGAAGAAGAGG-5′ (SEQ ID NO: 2219) AAT-492  Target: 5′-TCCAACAGCACCAATATCTTCTTCTCC-3′ (SEQ ID NO: 2359) 5′-CCAACAGCACCAAUAUCUUCUUCUCCC-3′ (SEQ ID NO: 2780) 3′-GGUUGUCGUGGUUAUAGAAGAAGAGGG-5′ (SEQ ID NO: 2220) AAT-493  Target: 5′-CCAACAGCACCAATATCTTCTTCTCCC-3′ (SEQ ID NO: 2360) 5′-CAACAGCACCAAUAUCUUCUUCUCCCC-3′ (SEQ ID NO: 2781) 3′-GUUGUCGUGGUUAUAGAAGAAGAGGGG-5′ (SEQ ID NO: 2221) AAT-494  Target: 5′-CAACAGCACCAATATCTTCTTCTCCCC-3′ (SEQ ID NO: 2361) 5′-AACAGCACCAAUAUCUUCUUCUCCCCA-3′ (SEQ ID NO: 2782) 3′-UUGUCGUGGUUAUAGAAGAAGAGGGGU-5′ (SEQ ID NO: 2222) AAT-495  Target: 5′-AACAGCACCAATATCTTCTTCTCCCCA-3′ (SEQ ID NO: 2362) 5′-ACAGCACCAAUAUCUUCUUCUCCCCAG-3′ (SEQ ID NO: 2783) 3′-UGUCGUGGUUAUAGAAGAAGAGGGGUC-5′ (SEQ ID NO: 2223) AAT-496  Target: 5′-ACAGCACCAATATCTTCTTCTCCCCAG-3′ (SEQ ID NO: 2363) 5′-CAGCACCAAUAUCUUCUUCUCCCCAGU-3′ (SEQ ID NO: 2784) 3′-GUCGUGGUUAUAGAAGAAGAGGGGUCA-5′ (SEQ ID NO: 2224) AAT-497  Target: 5′-CAGCACCAATATCTTCTTCTCCCCAGT-3′ (SEQ ID NO: 2364) 5′-AGCACCAAUAUCUUCUUCUCCCCAGUG-3′ (SEQ ID NO: 2785) 3′-UCGUGGUUAUAGAAGAAGAGGGGUCAC-5′ (SEQ ID NO: 2225) AAT-498  Target: 5′-AGCACCAATATCTTCTTCTCCCCAGTG-3′ (SEQ ID NO: 2365) 5′-GCACCAAUAUCUUCUUCUCCCCAGUGA-3′ (SEQ ID NO: 2786) 3′-CGUGGUUAUAGAAGAAGAGGGGUCACU-5′ (SEQ ID NO: 2226) AAT-499  Target: 5′-GCACCAATATCTTCTTCTCCCCAGTGA-3′ (SEQ ID NO: 2366) 5′-UCCCCAGUGAGCAUCGCUACAGCCUUU-3′ (SEQ ID NO: 2787) 3′-AGGGGUCACUCGUAGCGAUGUCGGAAA-5′ (SEQ ID NO: 2227) AAT-516  Target: 5′-TCCCCAGTGAGCATCGCTACAGCCTTT-3′ (SEQ ID NO: 2367) 5′-CCCCAGUGAGCAUCGCUACAGCCUUUG-3′ (SEQ ID NO: 2788) 3′-GGGGUCACUCGUAGCGAUGUCGGAAAC-5′ (SEQ ID NO: 2228) AAT-517  Target: 5′-CCCCAGTGAGCATCGCTACAGCCTTTG-3′ (SEQ ID NO: 2368) 5′-CCCAGUGAGCAUCGCUACAGCCUUUGC-3′ (SEQ ID NO: 2789) 3′-GGGUCACUCGUAGCGAUGUCGGAAACG-5′ (SEQ ID NO: 2229) AAT-518  Target: 5′-CCCAGTGAGCATCGCTACAGCCTTTGC-3′ (SEQ ID NO: 2369) 5′-CCAGUGAGCAUCGCUACAGCCUUUGCA-3′ (SEQ ID NO: 2790) 3′-GGUCACUCGUAGCGAUGUCGGAAACGU-5′ (SEQ ID NO: 2230) AAT-519  Target: 5′-CCAGTGAGCATCGCTACAGCCTTTGCA-3′ (SEQ ID NO: 2370) 5′-CAGUGAGCAUCGCUACAGCCUUUGCAA-3′ (SEQ ID NO: 2791) 3′-GUCACUCGUAGCGAUGUCGGAAACGUU-5′ (SEQ ID NO: 2231) AAT-520  Target: 5′-CAGTGAGCATCGCTACAGCCTTTGCAA-3′ (SEQ ID NO: 2371) 5′-AGUGAGCAUCGCUACAGCCUUUGCAAU-3′ (SEQ ID NO: 2792) 3′-UCACUCGUAGCGAUGUCGGAAACGUUA-5′ (SEQ ID NO: 2232) AAT-521  Target: 5′-AGTGAGCATCGCTACAGCCTTTGCAAT-3′ (SEQ ID NO: 2372) 5′-GUGAGCAUCGCUACAGCCUUUGCAAUG-3′ (SEQ ID NO: 2793) 3′-CACUCGUAGCGAUGUCGGAAACGUUAC-5′ (SEQ ID NO: 2233) AAT-522  Target: 5′-GTGAGCATCGCTACAGCCTTTGCAATG-3′ (SEQ ID NO: 2373) 5′-UGAGCAUCGCUACAGCCUUUGCAAUGC-3′ (SEQ ID NO: 2794) 3′-ACUCGUAGCGAUGUCGGAAACGUUACG-5′ (SEQ ID NO: 2234) AAT-523  Target: 5′-TGAGCATCGCTACAGCCTTTGCAATGC-3′ (SEQ ID NO: 2374) 5′-GAGCAUCGCUACAGCCUUUGCAAUGCU-3′ (SEQ ID NO: 2795) 3′-CUCGUAGCGAUGUCGGAAACGUUACGA-5′ (SEQ ID NO: 2235) AAT-524  Target: 5′-GAGCATCGCTACAGCCTTTGCAATGCT-3′ (SEQ ID NO: 2375) 5′-AGCAUCGCUACAGCCUUUGCAAUGCUC-3′ (SEQ ID NO: 2796) 3′-UCGUAGCGAUGUCGGAAACGUUACGAG-5′ (SEQ ID NO: 2236) AAT-525  Target: 5′-AGCATCGCTACAGCCTTTGCAATGCTC-3′ (SEQ ID NO: 2376) 5′-GCAUCGCUACAGCCUUUGCAAUGCUCU-3′ (SEQ ID NO: 2797) 3′-CGUAGCGAUGUCGGAAACGUUACGAGA-5′ (SEQ ID NO: 2237) AAT-526  Target: 5′-GCATCGCTACAGCCTTTGCAATGCTCT-3′ (SEQ ID NO: 2377) 5′-CAUCGCUACAGCCUUUGCAAUGCUCUC-3′ (SEQ ID NO: 2798) 3′-GUAGCGAUGUCGGAAACGUUACGAGAG-5′ (SEQ ID NO: 2238) AAT-527  Target: 5′-CATCGCTACAGCCTTTGCAATGCTCTC-3′ (SEQ ID NO: 2378) 5′-AUCGCUACAGCCUUUGCAAUGCUCUCC-3′ (SEQ ID NO: 2799) 3′-UAGCGAUGUCGGAAACGUUACGAGAGG-5′ (SEQ ID NO: 2239) AAT-528  Target: 5′-ATCGCTACAGCCTTTGCAATGCTCTCC-3′ (SEQ ID NO: 2379) 5′-UCGCUACAGCCUUUGCAAUGCUCUCCC-3′ (SEQ ID NO: 2800) 3′-AGCGAUGUCGGAAACGUUACGAGAGGG-5′ (SEQ ID NO: 2240) AAT-529  Target: 5′-TCGCTACAGCCTTTGCAATGCTCTCCC-3′ (SEQ ID NO: 2380) 5′-CGCUACAGCCUUUGCAAUGCUCUCCCU-3′ (SEQ ID NO: 2801) 3′-GCGAUGUCGGAAACGUUACGAGAGGGA-5′ (SEQ ID NO: 2241) AAT-530  Target: 5′-CGCTACAGCCTTTGCAATGCTCTCCCT-3′ (SEQ ID NO: 2381) 5′-GCUACAGCCUUUGCAAUGCUCUCCCUG-3′ (SEQ ID NO: 2802) 3′-CGAUGUCGGAAACGUUACGAGAGGGAC-5′ (SEQ ID NO: 2242) AAT-531  Target: 5′-GCTACAGCCTTTGCAATGCTCTCCCTG-3′ (SEQ ID NO: 2382) 5′-UCCCUGGGGACCAAGGCUGACACUCAC-3′ (SEQ ID NO: 2803) 3′-AGGGACCCCUGGUUCCGACUGUGAGUG-5′ (SEQ ID NO: 2243) AAT-552  Target: 5′-TCCCTGGGGACCAAGGCTGACACTCAC-3′ (SEQ ID NO: 2383) 5′-UGGGGACCAAGGCUGACACUCACGAUG-3′ (SEQ ID NO: 2804) 3′-ACCCCUGGUUCCGACUGUGAGUGCUAC-5′ (SEQ ID NO: 2244) AAT-556  Target: 5′-TGGGGACCAAGGCTGACACTCACGATG-3′ (SEQ ID NO: 2384) 5′-GGGGACCAAGGCUGACACUCACGAUGA-3′ (SEQ ID NO: 2805) 3′-CCCCUGGUUCCGACUGUGAGUGCUACU-5′ (SEQ ID NO: 2245) AAT-557  Target: 5′-GGGGACCAAGGCTGACACTCACGATGA-3′ (SEQ ID NO: 2385) 5′-GGGACCAAGGCUGACACUCACGAUGAA-3′ (SEQ ID NO: 2806) 3′-CCCUGGUUCCGACUGUGAGUGCUACUU-5′ (SEQ ID NO: 2246) AAT-558  Target: 5′-GGGACCAAGGCTGACACTCACGATGAA-3′ (SEQ ID NO: 2386) 5′-GAUGAAAUCCUGGAGGGCCUGAAUUUC-3′ (SEQ ID NO: 2807) 3′-CUACUUUAGGACCUCCCGGACUUAAAG-5′ (SEQ ID NO: 2247) AAT-579  Target: 5′-GATGAAATCCTGGAGGGCCTGAATTTC-3′ (SEQ ID NO: 2387) 5′-AUGAAAUCCUGGAGGGCCUGAAUUUCA-3′ (SEQ ID NO: 2808) 3′-UACUUUAGGACCUCCCGGACUUAAAGU-5′ (SEQ ID NO: 2248) AAT-580  Target: 5′-ATGAAATCCTGGAGGGCCTGAATTTCA-3′ (SEQ ID NO: 2388) 5′-GAUCCAUGAAGGCUUCCAGGAACUCCU-3′ (SEQ ID NO: 2809) 3′-CUAGGUACUUCCGAAGGUCCUUGAGGA-5′ (SEQ ID NO: 2249) AAT-632  Target: 5′-GATCCATGAAGGCTTCCAGGAACTCCT-3′ (SEQ ID NO: 2389) 5′-AUCCAUGAAGGCUUCCAGGAACUCCUC-3′ (SEQ ID NO: 2810) 3′-UAGGUACUUCCGAAGGUCCUUGAGGAG-5′ (SEQ ID NO: 2250) AAT-633  Target: 5′-ATCCATGAAGGCTTCCAGGAACTCCTC-3′ (SEQ ID NO: 2390) 5′-GGGGACACCGAAGAGGCCAAGAAACAG-3′ (SEQ ID NO: 2811) 3′-CCCCUGUGGCUUCUCCGGUUCUUUGUC-5′ (SEQ ID NO: 2251) AAT-801  Target: 5′-GGGGACACCGAAGAGGCCAAGAAACAG-3′ (SEQ ID NO: 2391) 5′-GGGACACCGAAGAGGCCAAGAAACAGA-3′ (SEQ ID NO: 2812) 3′-CCCUGUGGCUUCUCCGGUUCUUUGUCU-5′ (SEQ ID NO: 2252) AAT-802  Target: 5′-GGGACACCGAAGAGGCCAAGAAACAGA-3′ (SEQ ID NO: 2392) 5′-GGACACCGAAGAGGCCAAGAAACAGAU-3′ (SEQ ID NO: 2813) 3′-CCUGUGGCUUCUCCGGUUCUUUGUCUA-5′ (SEQ ID NO: 2253) AAT-803  Target: 5′-GGACACCGAAGAGGCCAAGAAACAGAT-3′ (SEQ ID NO: 2393) 5′-GACACCGAAGAGGCCAAGAAACAGAUC-3′ (SEQ ID NO: 2814) 3′-CUGUGGCUUCUCCGGUUCUUUGUCUAG-5′ (SEQ ID NO: 2254) AAT-804  Target: 5′-GACACCGAAGAGGCCAAGAAACAGATC-3′ (SEQ ID NO: 2394) 5′-ACACCGAAGAGGCCAAGAAACAGAUCA-3′ (SEQ ID NO: 2815) 3′-UGUGGCUUCUCCGGUUCUUUGUCUAGU-5′ (SEQ ID NO: 2255) AAT-805  Target: 5′-ACACCGAAGAGGCCAAGAAACAGATCA-3′ (SEQ ID NO: 2395) 5′-CACCGAAGAGGCCAAGAAACAGAUCAA-3′ (SEQ ID NO: 2816) 3′-GUGGCUUCUCCGGUUCUUUGUCUAGUU-5′ (SEQ ID NO: 2256) AAT-806  Target: 5′-CACCGAAGAGGCCAAGAAACAGATCAA-3′ (SEQ ID NO: 2396) 5′-ACCGAAGAGGCCAAGAAACAGAUCAAC-3′ (SEQ ID NO: 2817) 3′-UGGCUUCUCCGGUUCUUUGUCUAGUUG-5′ (SEQ ID NO: 2257) AAT-807  Target: 5′-ACCGAAGAGGCCAAGAAACAGATCAAC-3′ (SEQ ID NO: 2397) 5′-CCGAAGAGGCCAAGAAACAGAUCAACG-3′ (SEQ ID NO: 2818) 3′-GGCUUCUCCGGUUCUUUGUCUAGUUGC-5′ (SEQ ID NO: 2258) AAT-808  Target: 5′-CCGAAGAGGCCAAGAAACAGATCAACG-3′ (SEQ ID NO: 2398) 5′-CGAAGAGGCCAAGAAACAGAUCAACGA-3′ (SEQ ID NO: 2819) 3′-GCUUCUCCGGUUCUUUGUCUAGUUGCU-5′ (SEQ ID NO: 2259) AAT-809  Target: 5′-CGAAGAGGCCAAGAAACAGATCAACGA-3′ (SEQ ID NO: 2399) 5′-GAAGAGGCCAAGAAACAGAUCAACGAU-3′ (SEQ ID NO: 2820) 3′-CUUCUCCGGUUCUUUGUCUAGUUGCUA-5′ (SEQ ID NO: 2260) AAT-810  Target: 5′-GAAGAGGCCAAGAAACAGATCAACGAT-3′ (SEQ ID NO: 2400) 5′-AAGAGGCCAAGAAACAGAUCAACGAUU-3′ (SEQ ID NO: 2821) 3′-UUCUCCGGUUCUUUGUCUAGUUGCUAA-5′ (SEQ ID NO: 2261) AAT-811  Target: 5′-AAGAGGCCAAGAAACAGATCAACGATT-3′ (SEQ ID NO: 2401) 5′-AGAGGCCAAGAAACAGAUCAACGAUUA-3′ (SEQ ID NO: 2822) 3′-UCUCCGGUUCUUUGUCUAGUUGCUAAU-5′ (SEQ ID NO: 2262) AAT-812  Target: 5′-AGAGGCCAAGAAACAGATCAACGATTA-3′ (SEQ ID NO: 2402) 5′-GAGGCCAAGAAACAGAUCAACGAUUAC-3′ (SEQ ID NO: 2823) 3′-CUCCGGUUCUUUGUCUAGUUGCUAAUG-5′ (SEQ ID NO: 2263) AAT-813  Target: 5′-GAGGCCAAGAAACAGATCAACGATTAC-3′ (SEQ ID NO: 2403) 5′-GUUUUUGCUCUGGUGAAUUACAUCUUC-3′ (SEQ ID NO: 2824) 3′-CAAAAACGAGACCACUUAAUGUAGAAG-5′ (SEQ ID NO: 2264) AAT-900  Target: 5′-GTTTTTGCTCTGGTGAATTACATCTTC-3′ (SEQ ID NO: 2404) 5′-UUUUUGCUCUGGUGAAUUACAUCUUCU-3′ (SEQ ID NO: 2825) 3′-AAAAACGAGACCACUUAAUGUAGAAGA-5′ (SEQ ID NO: 2265) AAT-901  Target: 5′-TTTTTGCTCTGGTGAATTACATCTTCT-3′ (SEQ ID NO: 2405) 5′-UUUUGCUCUGGUGAAUUACAUCUUCUU-3′ (SEQ ID NO: 2826) 3′-AAAACGAGACCACUUAAUGUAGAAGAA-5′ (SEQ ID NO: 2266) AAT-902  Target: 5′-TTTTGCTCTGGTGAATTACATCTTCTT-3′ (SEQ ID NO: 2406) 5′-UUUGCUCUGGUGAAUUACAUCUUCUUU-3′ (SEQ ID NO: 2827) 3′-AAACGAGACCACUUAAUGUAGAAGAAA-5′ (SEQ ID NO: 2267) AAT-903  Target: 5′-TTTGCTCTGGTGAATTACATCTTCTTT-3′ (SEQ ID NO: 2407) 5′-UUGCUCUGGUGAAUUACAUCUUCUUUA-3′ (SEQ ID NO: 2828) 3′-AACGAGACCACUUAAUGUAGAAGAAAU-5′ (SEQ ID NO: 2268) AAT-904  Target: 5′-TTGCTCTGGTGAATTACATCTTCTTTA-3′ (SEQ ID NO: 2408) 5′-UGCUCUGGUGAAUUACAUCUUCUUUAA-3′ (SEQ ID NO: 2829) 3′-ACGAGACCACUUAAUGUAGAAGAAAUU-5′ (SEQ ID NO: 2269) AAT-905  Target: 5′-TGCTCTGGTGAATTACATCTTCTTTAA-3′ (SEQ ID NO: 2409) 5′-GCUCUGGUGAAUUACAUCUUCUUUAAA-3′ (SEQ ID NO: 2830) 3′-CGAGACCACUUAAUGUAGAAGAAAUUU-5′ (SEQ ID NO: 2270) AAT-906  Target: 5′-GCTCTGGTGAATTACATCTTCTTTAAA-3′ (SEQ ID NO: 2410) 5′-CUCUGGUGAAUUACAUCUUCUUUAAAG-3′ (SEQ ID NO: 2831) 3′-GAGACCACUUAAUGUAGAAGAAAUUUC-5′ (SEQ ID NO: 2271) AAT-907  Target: 5′-CTCTGGTGAATTACATCTTCTTTAAAG-3′ (SEQ ID NO: 2411) 5′-UCUGGUGAAUUACAUCUUCUUUAAAGG-3′ (SEQ ID NO: 2832) 3′-AGACCACUUAAUGUAGAAGAAAUUUCC-5′ (SEQ ID NO: 2272) AAT-908  Target: 5′-TCTGGTGAATTACATCTTCTTTAAAGG-3′ (SEQ ID NO: 2412) 5′-CUGGUGAAUUACAUCUUCUUUAAAGGC-3′ (SEQ ID NO: 2833) 3′-GACCACUUAAUGUAGAAGAAAUUUCCG-5′ (SEQ ID NO: 2273) AAT-909  Target: 5′-CTGGTGAATTACATCTTCTTTAAAGGC-3′ (SEQ ID NO: 2413) 5′-UGGUGAAUUACAUCUUCUUUAAAGGCA-3′ (SEQ ID NO: 2834) 3′-ACCACUUAAUGUAGAAGAAAUUUCCGU-5′ (SEQ ID NO: 2274) AAT-910  Target: 5′-TGGTGAATTACATCTTCTTTAAAGGCA-3′ (SEQ ID NO: 2414) 5′-GGUGAAUUACAUCUUCUUUAAAGGCAA-3′ (SEQ ID NO: 2835) 3′-CCACUUAAUGUAGAAGAAAUUUCCGUU-5′ (SEQ ID NO: 2275) AAT-911  Target: 5′-GGTGAATTACATCTTCTTTAAAGGCAA-3′ (SEQ ID NO: 2415) 5′-GUGAAUUACAUCUUCUUUAAAGGCAAA-3′ (SEQ ID NO: 2836) 3′-CACUUAAUGUAGAAGAAAUUUCCGUUU-5′ (SEQ ID NO: 2276) AAT-912  Target: 5′-GTGAATTACATCTTCTTTAAAGGCAAA-3′ (SEQ ID NO: 2416) 5′-UGAAUUACAUCUUCUUUAAAGGCAAAU-3′ (SEQ ID NO: 2837) 3′-ACUUAAUGUAGAAGAAAUUUCCGUUUA-5′ (SEQ ID NO: 2277) AAT-913  Target: 5′-TGAATTACATCTTCTTTAAAGGCAAAT-3′ (SEQ ID NO: 2417) 5′-GAAUUACAUCUUCUUUAAAGGCAAAUG-3′ (SEQ ID NO: 2838) 3′-CUUAAUGUAGAAGAAAUUUCCGUUUAC-5′ (SEQ ID NO: 2278) AAT-914  Target: 5′-GAATTACATCTTCTTTAAAGGCAAATG-3′ (SEQ ID NO: 2418) 5′-AAUUACAUCUUCUUUAAAGGCAAAUGG-3′ (SEQ ID NO: 2839) 3′-UUAAUGUAGAAGAAAUUUCCGUUUACC-5′ (SEQ ID NO: 2279) AAT-915  Target: 5′-AATTACATCTTCTTTAAAGGCAAATGG-3′ (SEQ ID NO: 2419) 5′-AUUACAUCUUCUUUAAAGGCAAAUGGG-3′ (SEQ ID NO: 2840) 3′-UAAUGUAGAAGAAAUUUCCGUUUACCC-5′ (SEQ ID NO: 2280) AAT-916  Target: 5′-ATTACATCTTCTTTAAAGGCAAATGGG-3′ (SEQ ID NO: 2420) 5′-UUACAUCUUCUUUAAAGGCAAAUGGGA-3′ (SEQ ID NO: 2841) 3′-AAUGUAGAAGAAAUUUCCGUUUACCCU-5′ (SEQ ID NO: 2281) AAT-917  Target: 5′-TTACATCTTCTTTAAAGGCAAATGGGA-3′ (SEQ ID NO: 2421) 5′-UACAUCUUCUUUAAAGGCAAAUGGGAG-3′ (SEQ ID NO: 2842) 3′-AUGUAGAAGAAAUUUCCGUUUACCCUC-5′ (SEQ ID NO: 2282) AAT-918  Target: 5′-TACATCTTCTTTAAAGGCAAATGGGAG-3′ (SEQ ID NO: 2422) 5′-UCUUCUUUAAAGGCAAAUGGGAGAGAC-3′ (SEQ ID NO: 2843) 3′-AGAAGAAAUUUCCGUUUACCCUCUCUG-5′ (SEQ ID NO: 2283) AAT-922  Target: 5′-TCTTCTTTAAAGGCAAATGGGAGAGAC-3′ (SEQ ID NO: 2423) 5′-UUCUUUAAAGGCAAAUGGGAGAGACCC-3′ (SEQ ID NO: 2844) 3′-AAGAAAUUUCCGUUUACCCUCUCUGGG-5′ (SEQ ID NO: 2284) AAT-924  Target: 5′-TTCTTTAAAGGCAAATGGGAGAGACCC-3′ (SEQ ID NO: 2424) 5′-AGGCAAAUGGGAGAGACCCUUUGAAGU-3′ (SEQ ID NO: 2845) 3′-UCCGUUUACCCUCUCUGGGAAACUUCA-5′ (SEQ ID NO: 2285) AAT-932  Target: 5′-AGGCAAATGGGAGAGACCCTTTGAAGT-3′ (SEQ ID NO: 2425) 5′-GGCAAAUGGGAGAGACCCUUUGAAGUC-3′ (SEQ ID NO: 2846) 3′-CCGUUUACCCUCUCUGGGAAACUUCAG-5′ (SEQ ID NO: 2286) AAT-933  Target: 5′-GGCAAATGGGAGAGACCCTTTGAAGTC-3′ (SEQ ID NO: 2426) 5′-GCAAAUGGGAGAGACCCUUUGAAGUCA-3′ (SEQ ID NO: 2847) 3′-CGUUUACCCUCUCUGGGAAACUUCAGU-5′ (SEQ ID NO: 2287) AAT-934  Target: 5′-GCAAATGGGAGAGACCCTTTGAAGTCA-3′ (SEQ ID NO: 2427) 5′-CAAAUGGGAGAGACCCUUUGAAGUCAA-3′ (SEQ ID NO: 2848) 3′-GUUUACCCUCUCUGGGAAACUUCAGUU-5′ (SEQ ID NO: 2288) AAT-935  Target: 5′-CAAATGGGAGAGACCCTTTGAAGTCAA-3′ (SEQ ID NO: 2428) 5′-GCUGUCCAGCUGGGUGCUGCUGAUGAA-3′ (SEQ ID NO: 2849) 3′-CGACAGGUCGACCCACGACGACUACUU-5′ (SEQ ID NO: 2289) AAT-1061 Target: 5′-GCTGTCCAGCTGGGTGCTGCTGATGAA-3′ (SEQ ID NO: 2429) 5′-CUGUCCAGCUGGGUGCUGCUGAUGAAA-3′ (SEQ ID NO: 2850) 3′-GACAGGUCGACCCACGACGACUACUUU-5′ (SEQ ID NO: 2290) AAT-1062 Target: 5′-CTGTCCAGCTGGGTGCTGCTGATGAAA-3′ (SEQ ID NO: 2430) 5′-UGUCCAGCUGGGUGCUGCUGAUGAAAU-3′ (SEQ ID NO: 2851) 3′-ACAGGUCGACCCACGACGACUACUUUA-5′ (SEQ ID NO: 2291) AAT-1063 Target: 5′-TGTCCAGCTGGGTGCTGCTGATGAAAT-3′ (SEQ ID NO: 2431) 5′-GUCCAGCUGGGUGCUGCUGAUGAAAUA-3′ (SEQ ID NO: 2852) 3′-CAGGUCGACCCACGACGACUACUUUAU-5′ (SEQ ID NO: 2292) AAT-1064 Target: 5′-GTCCAGCTGGGTGCTGCTGATGAAATA-3′ (SEQ ID NO: 2432) 5′-UCCAGCUGGGUGCUGCUGAUGAAAUAC-3′ (SEQ ID NO: 2853) 3′-AGGUCGACCCACGACGACUACUUUAUG-5′ (SEQ ID NO: 2293) AAT-1065 Target: 5′-TCCAGCTGGGTGCTGCTGATGAAATAC-3′ (SEQ ID NO: 2433) 5′-CCAGCUGGGUGCUGCUGAUGAAAUACC-3′ (SEQ ID NO: 2854) 3′-GGUCGACCCACGACGACUACUUUAUGG-5′ (SEQ ID NO: 2294) AAT-1066 Target: 5′-CCAGCTGGGTGCTGCTGATGAAATACC-3′ (SEQ ID NO: 2434) 5′-CAGCUGGGUGCUGCUGAUGAAAUACCU-3′ (SEQ ID NO: 2855) 3′-GUCGACCCACGACGACUACUUUAUGGA-5′ (SEQ ID NO: 2295) AAT-1067 Target: 5′-CAGCTGGGTGCTGCTGATGAAATACCT-3′ (SEQ ID NO: 2435) 5′-AGCUGGGUGCUGCUGAUGAAAUACCUG-3′ (SEQ ID NO: 2856) 3′-UCGACCCACGACGACUACUUUAUGGAC-5′ (SEQ ID NO: 2296) AAT-1068 Target: 5′-AGCTGGGTGCTGCTGATGAAATACCTG-3′ (SEQ ID NO: 2436) 5′-GCUGGGUGCUGCUGAUGAAAUACCUGG-3′ (SEQ ID NO: 2857) 3′-CGACCCACGACGACUACUUUAUGGACC-5′ (SEQ ID NO: 2297) AAT-1069 Target: 5′-GCTGGGTGCTGCTGATGAAATACCTGG-3′ (SEQ ID NO: 2437) 5′-CUGGGUGCUGCUGAUGAAAUACCUGGG-3′ (SEQ ID NO: 2858) 3′-GACCCACGACGACUACUUUAUGGACCC-5′ (SEQ ID NO: 2298) AAT-1070 Target: 5′-CTGGGTGCTGCTGATGAAATACCTGGG-3′ (SEQ ID NO: 2438) 5′-GGGUGCUGCUGAUGAAAUACCUGGGCA-3′ (SEQ ID NO: 2859) 3′-CCCACGACGACUACUUUAUGGACCCGU-5′ (SEQ ID NO: 2299) AAT-1072 Target: 5′-GGGTGCTGCTGATGAAATACCTGGGCA-3′ (SEQ ID NO: 2439) 5′-GGUGCUGCUGAUGAAAUACCUGGGCAA-3′ (SEQ ID NO: 2860) 3′-CCACGACGACUACUUUAUGGACCCGUU-5′ (SEQ ID NO: 2300) AAT-1073 Target: 5′-GGTGCTGCTGATGAAATACCTGGGCAA-3′ (SEQ ID NO: 2440) 5′-GUGCUGCUGAUGAAAUACCUGGGCAAU-3′ (SEQ ID NO: 2861) 3′-CACGACGACUACUUUAUGGACCCGUUA-5′ (SEQ ID NO: 2301) AAT-1074 Target: 5′-GTGCTGCTGATGAAATACCTGGGCAAT-3′ (SEQ ID NO: 2441) 5′-UGCUGCUGAUGAAAUACCUGGGCAAUG-3′ (SEQ ID NO: 2862) 3′-ACGACGACUACUUUAUGGACCCGUUAC-5′ (SEQ ID NO: 2302) AAT-1075 Target: 5′-TGCTGCTGATGAAATACCTGGGCAATG-3′ (SEQ ID NO: 2442) 5′-GCUGCUGAUGAAAUACCUGGGCAAUGC-3′ (SEQ ID NO: 2863) 3′-CGACGACUACUUUAUGGACCCGUUACG-5′ (SEQ ID NO: 2303) AAT-1076 Target: 5′-GCTGCTGATGAAATACCTGGGCAATGC-3′ (SEQ ID NO: 2443) 5′-CUGCUGAUGAAAUACCUGGGCAAUGCC-3′ (SEQ ID NO: 2864) 3′-GACGACUACUUUAUGGACCCGUUACGG-5′ (SEQ ID NO: 2304) AAT-1077 Target: 5′-CTGCTGATGAAATACCTGGGCAATGCC-3′ (SEQ ID NO: 2444) 5′-UGCUGAUGAAAUACCUGGGCAAUGCCA-3′ (SEQ ID NO: 2865) 3′-ACGACUACUUUAUGGACCCGUUACGGU-5′ (SEQ ID NO: 2305) AAT-1078 Target: 5′-TGCTGATGAAATACCTGGGCAATGCCA-3′ (SEQ ID NO: 2445) 5′-GCUGAUGAAAUACCUGGGCAAUGCCAC-3′ (SEQ ID NO: 2866) 3′-CGACUACUUUAUGGACCCGUUACGGUG-5′ (SEQ ID NO: 2306) AAT-1079 Target: 5′-GCTGATGAAATACCTGGGCAATGCCAC-3′ (SEQ ID NO: 2446) 5′-CUGAUGAAAUACCUGGGCAAUGCCACC-3′ (SEQ ID NO: 2867) 3′-GACUACUUUAUGGACCCGUUACGGUGG-5′ (SEQ ID NO: 2307) AAT-1080 Target: 5′-CTGATGAAATACCTGGGCAATGCCACC-3′ (SEQ ID NO: 2447) 5′-UGAUGAAAUACCUGGGCAAUGCCACCG-3′ (SEQ ID NO: 2868) 3′-ACUACUUUAUGGACCCGUUACGGUGGC-5′ (SEQ ID NO: 2308) AAT-1081 Target: 5′-TGATGAAATACCTGGGCAATGCCACCG-3′ (SEQ ID NO: 2448) 5′-AUGAAAUACCUGGGCAAUGCCACCGCC-3′ (SEQ ID NO: 2869) 3′-UACUUUAUGGACCCGUUACGGUGGCGG-5′ (SEQ ID NO: 2309) AAT-1083 Target: 5′-ATGAAATACCTGGGCAATGCCACCGCC-3′ (SEQ ID NO: 2449) 5′-UACAGCACCUGGAAAAUGAACUCACCC-3′ (SEQ ID NO: 2870) 3′-AUGUCGUGGACCUUUUACUUGAGUGGG-5′ (SEQ ID NO: 2310) AAT-1138 Target: 5′-TACAGCACCTGGAAAATGAACTCACCC-3′ (SEQ ID NO: 2450) 5′-ACCUGGAAAAUGAACUCACCCACGAUA-3′ (SEQ ID NO: 2871) 3′-UGGACCUUUUACUUGAGUGGGUGCUAU-5′ (SEQ ID NO: 2311) AAT-1144 Target: 5′-ACCTGGAAAATGAACTCACCCACGATA-3′ (SEQ ID NO: 2451) 5′-CCUGGAAAAUGAACUCACCCACGAUAU-3′ (SEQ ID NO: 2872) 3′-GGACCUUUUACUUGAGUGGGUGCUAUA-5′ (SEQ ID NO: 2312) AAT-1145 Target: 5′-CCTGGAAAATGAACTCACCCACGATAT-3′ (SEQ ID NO: 2452) 5′-ACGAUAUCAUCACCAAGUUCCUGGAAA-3′ (SEQ ID NO: 2873) 3′-UGCUAUAGUAGUGGUUCAAGGACCUUU-5′ (SEQ ID NO: 2313) AAT-1165 Target: 5′-ACGATATCATCACCAAGTTCCTGGAAA-3′ (SEQ ID NO: 2453) 5′-ACCAAGUUCCUGGAAAAUGAAGACAGA-3′ (SEQ ID NO: 2874) 3′-UGGUUCAAGGACCUUUUACUUCUGUCU-5′ (SEQ ID NO: 2314) AAT-1176 Target: 5′-ACCAAGTTCCTGGAAAATGAAGACAGA-3′ (SEQ ID NO: 2454) 5′-GUCCAUUACUGGAACCUAUGAUCUGAA-3′ (SEQ ID NO: 2875) 3′-CAGGUAAUGACCUUGGAUACUAGACUU-5′ (SEQ ID NO: 2315) AAT-1232 Target: 5′-GTCCATTACTGGAACCTATGATCTGAA-3′ (SEQ ID NO: 2455) 5′-UCCAUUACUGGAACCUAUGAUCUGAAG-3′ (SEQ ID NO: 2876) 3′-AGGUAAUGACCUUGGAUACUAGACUUC-5′ (SEQ ID NO: 2316) AAT-1233 Target: 5′-TCCATTACTGGAACCTATGATCTGAAG-3′ (SEQ ID NO: 2456) 5′-CCAUUACUGGAACCUAUGAUCUGAAGA-3′ (SEQ ID NO: 2877) 3′-GGUAAUGACCUUGGAUACUAGACUUCU-5′ (SEQ ID NO: 2317) AAT-1234 Target: 5′-CCATTACTGGAACCTATGATCTGAAGA-3′ (SEQ ID NO: 2457) 5′-CAUUACUGGAACCUAUGAUCUGAAGAG-3′ (SEQ ID NO: 2878) 3′-GUAAUGACCUUGGAUACUAGACUUCUC-5′ (SEQ ID NO: 2318) AAT-1235 Target: 5′-CATTACTGGAACCTATGATCTGAAGAG-3′ (SEQ ID NO: 2458) 5′-AUUACUGGAACCUAUGAUCUGAAGAGC-3′ (SEQ ID NO: 2879) 3′-UAAUGACCUUGGAUACUAGACUUCUCG-5′ (SEQ ID NO: 2319) AAT-1236 Target: 5′-ATTACTGGAACCTATGATCTGAAGAGC-3′ (SEQ ID NO: 2459) 5′-UUACUGGAACCUAUGAUCUGAAGAGCG-3′ (SEQ ID NO: 2880) 3′-AAUGACCUUGGAUACUAGACUUCUCGC-5′ (SEQ ID NO: 2320) AAT-1237 Target: 5′-TTACTGGAACCTATGATCTGAAGAGCG-3′ (SEQ ID NO: 2460) 5′-UACUGGAACCUAUGAUCUGAAGAGCGU-3′ (SEQ ID NO: 2881) 3′-AUGACCUUGGAUACUAGACUUCUCGCA-5′ (SEQ ID NO: 2321) AAT-1238 Target: 5′-TACTGGAACCTATGATCTGAAGAGCGT-3′ (SEQ ID NO: 2461) 5′-ACUGGAACCUAUGAUCUGAAGAGCGUC-3′ (SEQ ID NO: 2882) 3′-UGACCUUGGAUACUAGACUUCUCGCAG-5′ (SEQ ID NO: 2322) AAT-1239 Target: 5′-ACTGGAACCTATGATCTGAAGAGCGTC-3′ (SEQ ID NO: 2462) 5′-CUGGAACCUAUGAUCUGAAGAGCGUCC-3′ (SEQ ID NO: 2883) 3′-GACCUUGGAUACUAGACUUCUCGCAGG-5′ (SEQ ID NO: 2323) AAT-1240 Target: 5′-CTGGAACCTATGATCTGAAGAGCGTCC-3′ (SEQ ID NO: 2463) 5′-GCAUCACUAAGGUCUUCAGCAAUGGGG-3′ (SEQ ID NO: 2884) 3′-CGUAGUGAUUCCAGAAGUCGUUACCCC-5′ (SEQ ID NO: 2324) AAT-1279 Target: 5′-GCATCACTAAGGTCTTCAGCAATGGGG-3′ (SEQ ID NO: 2464) 5′-CAUCACUAAGGUCUUCAGCAAUGGGGC-3′ (SEQ ID NO: 2885) 3′-GUAGUGAUUCCAGAAGUCGUUACCCCG-5′ (SEQ ID NO: 2325) AAT-1280 Target: 5′-CATCACTAAGGTCTTCAGCAATGGGGC-3′ (SEQ ID NO: 2465) 5′-AUCACUAAGGUCUUCAGCAAUGGGGCU-3′ (SEQ ID NO: 2886) 3′-UAGUGAUUCCAGAAGUCGUUACCCCGA-5′ (SEQ ID NO: 2326) AAT-1281 Target: 5′-ATCACTAAGGTCTTCAGCAATGGGGCT-3′ (SEQ ID NO: 2466) 5′-CACUAAGGUCUUCAGCAAUGGGGCUGA-3′ (SEQ ID NO: 2887) 3′-GUGAUUCCAGAAGUCGUUACCCCGACU-5′ (SEQ ID NO: 2327) AAT-1283 Target: 5′-CACTAAGGTCTTCAGCAATGGGGCTGA-3′ (SEQ ID NO: 2467) 5′-ACUAAGGUCUUCAGCAAUGGGGCUGAC-3′ (SEQ ID NO: 2888) 3′-UGAUUCCAGAAGUCGUUACCCCGACUG-5′ (SEQ ID NO: 2328) AAT-1284 Target: 5′-ACTAAGGTCTTCAGCAATGGGGCTGAC-3′ (SEQ ID NO: 2468) 5′-CCUGAAGCUCUCCAAGGCCGUGCAUAA-3′ (SEQ ID NO: 2889) 3′-GGACUUCGAGAGGUUCCGGCACGUAUU-5′ (SEQ ID NO: 2329) AAT-1337 Target: 5′-CCTGAAGCTCTCCAAGGCCGTGCATAA-3′ (SEQ ID NO: 2469) 5′-CUGAAGCUCUCCAAGGCCGUGCAUAAG-3′ (SEQ ID NO: 2890) 3′-GACUUCGAGAGGUUCCGGCACGUAUUC-5′ (SEQ ID NO: 2330) AAT-1338 Target: 5′-CTGAAGCTCTCCAAGGCCGTGCATAAG-3′ (SEQ ID NO: 2470) 5′-UGAAGCUCUCCAAGGCCGUGCAUAAGG-3′ (SEQ ID NO: 2891) 3′-ACUUCGAGAGGUUCCGGCACGUAUUCC-5′ (SEQ ID NO: 2331) AAT-1339 Target: 5′-TGAAGCTCTCCAAGGCCGTGCATAAGG-3′ (SEQ ID NO: 2471) 5′-CCCCGAGGUCAAGUUCAACAAACCCUU-3′ (SEQ ID NO: 2892) 3′-GGGGCUCCAGUUCAAGUUGUUUGGGAA-5′ (SEQ ID NO: 2332) AAT-1442 Target: 5′-CCCCGAGGTCAAGTTCAACAAACCCTT-3′ (SEQ ID NO: 2472) 5′-CCCGAGGUCAAGUUCAACAAACCCUUU-3′ (SEQ ID NO: 2893) 3′-GGGCUCCAGUUCAAGUUGUUUGGGAAA-5′ (SEQ ID NO: 2333) AAT-1443 Target: 5′-CCCGAGGTCAAGTTCAACAAACCCTTT-3′ (SEQ ID NO: 2473) 5′-CCGAGGUCAAGUUCAACAAACCCUUUG-3′ (SEQ ID NO: 2894) 3′-GGCUCCAGUUCAAGUUGUUUGGGAAAC-5′ (SEQ ID NO: 2334) AAT-1444 Target: 5′-CCGAGGTCAAGTTCAACAAACCCTTTG-3′ (SEQ ID NO: 2474) 5′-CGAGGUCAAGUUCAACAAACCCUUUGU-3′ (SEQ ID NO: 2895) 3′-GCUCCAGUUCAAGUUGUUUGGGAAACA-5′ (SEQ ID NO: 2335) AAT-1445 Target: 5′-CGAGGTCAAGTTCAACAAACCCTTTGT-3′ (SEQ ID NO: 2475) 5′-GAGGUCAAGUUCAACAAACCCUUUGUC-3′ (SEQ ID NO: 2896) 3′-CUCCAGUUCAAGUUGUUUGGGAAACAG-5′ (SEQ ID NO: 2336) AAT-1446 Target: 5′-GAGGTCAAGTTCAACAAACCCTTTGTC-3′ (SEQ ID NO: 2476) 5′-AGGUCAAGUUCAACAAACCCUUUGUCU-3′ (SEQ ID NO: 2897) 3′-UCCAGUUCAAGUUGUUUGGGAAACAGA-5′ (SEQ ID NO: 2337) AAT-1447 Target: 5′-AGGTCAAGTTCAACAAACCCTTTGTCT-3′ (SEQ ID NO: 2477) 5′-GGUCAAGUUCAACAAACCCUUUGUCUU-3′ (SEQ ID NO: 2898) 3′-CCAGUUCAAGUUGUUUGGGAAACAGAA-5′ (SEQ ID NO: 2338) AAT-1448 Target: 5′-GGTCAAGTTCAACAAACCCTTTGTCTT-3′ (SEQ ID NO: 2478) 5′-GUCAAGUUCAACAAACCCUUUGUCUUC-3′ (SEQ ID NO: 2899) 3′-CAGUUCAAGUUGUUUGGGAAACAGAAG-5′ (SEQ ID NO: 2339) AAT-1449 Target: 5′-GTCAAGTTCAACAAACCCTTTGTCTTC-3′ (SEQ ID NO: 2479) 5′-UCAAGUUCAACAAACCCUUUGUCUUCU-3′ (SEQ ID NO: 2900) 3′-AGUUCAAGUUGUUUGGGAAACAGAAGA-5′ (SEQ ID NO: 2340) AAT-1450 Target: 5′-TCAAGTTCAACAAACCCTTTGTCTTCT-3′ (SEQ ID NO: 2480) 5′-CAAGUUCAACAAACCCUUUGUCUUCUU-3′ (SEQ ID NO: 2901) 3′-GUUCAAGUUGUUUGGGAAACAGAAGAA-5′ (SEQ ID NO: 2341) AAT-1451 Target: 5′-CAAGTTCAACAAACCCTTTGTCTTCTT-3′ (SEQ ID NO: 2481)

TABLE 16 Further DsiRNA Component 19 NucleotideTargetSequences in α-1 antitrypsin mRNA AAT-366 19 nt Target #1: 5′-CCAGGGAGAUGCUGCCCAG-3′ (SEQ ID NO: 2902) AAT-366 19 nt Target #2: 5′-CCCAGGGAGAUGCUGCCCA-3′ (SEQ ID NO: 3042) AAT-366 19 nt Target #3: 5′-CCCCAGGGAGAUGCUGCCC-3′ (SEQ ID NO: 3182) AAT-367 19 nt Target #1: 5′-CAGGGAGAUGCUGCCCAGA-3′ (SEQ ID NO: 2903) AAT-367 19 nt Target #2: 5′-CCAGGGAGAUGCUGCCCAG-3′ (SEQ ID NO: 3043) AAT-367 19 nt Target #3: 5′-CCCAGGGAGAUGCUGCCCA-3′ (SEQ ID NO: 3183) AAT-368 19 nt Target #1: 5′-AGGGAGAUGCUGCCCAGAA-3′ (SEQ ID NO: 2904) AAT-368 19 nt Target #2: 5′-CAGGGAGAUGCUGCCCAGA-3′ (SEQ ID NO: 3044) AAT-368 19 nt Target #3: 5′-CCAGGGAGAUGCUGCCCAG-3′ (SEQ ID NO: 3184) AAT-369 19 nt Target #1: 5′-GGGAGAUGCUGCCCAGAAG-3′ (SEQ ID NO: 2905) AAT-369 19 nt Target #2: 5′-AGGGAGAUGCUGCCCAGAA-3′ (SEQ ID NO: 3045) AAT-369 19 nt Target #3: 5′-CAGGGAGAUGCUGCCCAGA-3′ (SEQ ID NO: 3185) AAT-370 19 nt Target #1: 5′-GGAGAUGCUGCCCAGAAGA-3′ (SEQ ID NO: 2906) AAT-370 19 nt Target #2: 5′-GGGAGAUGCUGCCCAGAAG-3′ (SEQ ID NO: 3046) AAT-370 19 nt Target #3: 5′-AGGGAGAUGCUGCCCAGAA-3′ (SEQ ID NO: 3186) AAT-371 19 nt Target #1: 5′-GAGAUGCUGCCCAGAAGAC-3′ (SEQ ID NO: 2907) AAT-371 19 nt Target #2: 5′-GGAGAUGCUGCCCAGAAGA-3′ (SEQ ID NO: 3047) AAT-371 19 nt Target #3: 5′-GGGAGAUGCUGCCCAGAAG-3′ (SEQ ID NO: 3187) AAT-391 19 nt Target #1: 5′-GAUACAUCCCACCAUGAUC-3′ (SEQ ID NO: 2908) AAT-391 19 nt Target #2: 5′-AGAUACAUCCCACCAUGAU-3′ (SEQ ID NO: 3048) AAT-391 19 nt Target #3: 5′-CAGAUACAUCCCACCAUGA-3′ (SEQ ID NO: 3188) AAT-392 19 nt Target #1: 5′-AUACAUCCCACCAUGAUCA-3′ (SEQ ID NO: 2909) AAT-392 19 nt Target #2: 5′-GAUACAUCCCACCAUGAUC-3′ (SEQ ID NO: 3049) AAT-392 19 nt Target #3: 5′-AGAUACAUCCCACCAUGAU-3′ (SEQ ID NO: 3189) AAT-393 19 nt Target #1: 5′-UACAUCCCACCAUGAUCAG-3′ (SEQ ID NO: 2910) AAT-393 19 nt Target #2: 5′-AUACAUCCCACCAUGAUCA-3′ (SEQ ID NO: 3050) AAT-393 19 nt Target #3: 5′-GAUACAUCCCACCAUGAUC-3′ (SEQ ID NO: 3190) AAT-394 19 nt Target #1: 5′-ACAUCCCACCAUGAUCAGG-3′ (SEQ ID NO: 2911) AAT-394 19 nt Target #2: 5′-UACAUCCCACCAUGAUCAG-3′ (SEQ ID NO: 3051) AAT-394 19 nt Target #3: 5′-AUACAUCCCACCAUGAUCA-3′ (SEQ ID NO: 3191) AAT-485 19 nt Target #1: 5′-ACCAGUCCAACAGCACCAA-3′ (SEQ ID NO: 2912) AAT-485 19 nt Target #2: 5′-CACCAGUCCAACAGCACCA-3′ (SEQ ID NO: 3052) AAT-485 19 nt Target #3: 5′-ACACCAGUCCAACAGCACC-3′ (SEQ ID NO: 3192) AAT-486 19 nt Target #1: 5′-CCAGUCCAACAGCACCAAU-3′ (SEQ ID NO: 2913) AAT-486 19 nt Target #2: 5′-ACCAGUCCAACAGCACCAA-3′ (SEQ ID NO: 3053) AAT-486 19 nt Target #3: 5′-CACCAGUCCAACAGCACCA-3′ (SEQ ID NO: 3193) AAT-487 19 nt Target #1: 5′-CAGUCCAACAGCACCAAUA-3′ (SEQ ID NO: 2914) AAT-487 19 nt Target #2: 5′-CCAGUCCAACAGCACCAAU-3′ (SEQ ID NO: 3054) AAT-487 19 nt Target #3: 5′-ACCAGUCCAACAGCACCAA-3′ (SEQ ID NO: 3194) AAT-488 19 nt Target #1: 5′-AGUCCAACAGCACCAAUAU-3′ (SEQ ID NO: 2915) AAT-488 19 nt Target #2: 5′-CAGUCCAACAGCACCAAUA-3′ (SEQ ID NO: 3055) AAT-488 19 nt Target #3: 5′-CCAGUCCAACAGCACCAAU-3′ (SEQ ID NO: 3195) AAT-489 19 nt Target #1: 5′-GUCCAACAGCACCAAUAUC-3′ (SEQ ID NO: 2916) AAT-489 19 nt Target #2: 5′-AGUCCAACAGCACCAAUAU-3′ (SEQ ID NO: 3056) AAT-489 19 nt Target #3: 5′-CAGUCCAACAGCACCAAUA-3′ (SEQ ID NO: 3196) AAT-490 19 nt Target #1: 5′-UCCAACAGCACCAAUAUCU-3′ (SEQ ID NO: 2917) AAT-490 19 nt Target #2: 5′-GUCCAACAGCACCAAUAUC-3′ (SEQ ID NO: 3057) AAT-490 19 nt Target #3: 5′-AGUCCAACAGCACCAAUAU-3′ (SEQ ID NO: 3197) AAT-491 19 nt Target #1: 5′-CCAACAGCACCAAUAUCUU-3′ (SEQ ID NO: 2918) AAT-491 19 nt Target #2: 5′-UCCAACAGCACCAAUAUCU-3′ (SEQ ID NO: 3058) AAT-491 19 nt Target #3: 5′-GUCCAACAGCACCAAUAUC-3′ (SEQ ID NO: 3198) AAT-492 19 nt Target #1: 5′-CAACAGCACCAAUAUCUUC-3′ (SEQ ID NO: 2919) AAT-492 19 nt Target #2: 5′-CCAACAGCACCAAUAUCUU-3′ (SEQ ID NO: 3059) AAT-492 19 nt Target #3: 5′-UCCAACAGCACCAAUAUCU-3′ (SEQ ID NO: 3199) AAT-493 19 nt Target #1: 5′-AACAGCACCAAUAUCUUCU-3′ (SEQ ID NO: 2920) AAT-493 19 nt Target #2: 5′-CAACAGCACCAAUAUCUUC-3′ (SEQ ID NO: 3060) AAT-493 19 nt Target #3: 5′-CCAACAGCACCAAUAUCUU-3′ (SEQ ID NO: 3200) AAT-494 19 nt Target #1: 5′-ACAGCACCAAUAUCUUCUU-3′ (SEQ ID NO: 2921) AAT-494 19 nt Target #2: 5′-AACAGCACCAAUAUCUUCU-3′ (SEQ ID NO: 3061) AAT-494 19 nt Target #3: 5′-CAACAGCACCAAUAUCUUC-3′ (SEQ ID NO: 3201) AAT-495 19 nt Target #1: 5′-CAGCACCAAUAUCUUCUUC-3′ (SEQ ID NO: 2922) AAT-495 19 nt Target #2: 5′-ACAGCACCAAUAUCUUCUU-3′ (SEQ ID NO: 3062) AAT-495 19 nt Target #3: 5′-AACAGCACCAAUAUCUUCU-3′ (SEQ ID NO: 3202) AAT-496 19 nt Target #1: 5′-AGCACCAAUAUCUUCUUCU-3′ (SEQ ID NO: 2923) AAT-496 19 nt Target #2: 5′-CAGCACCAAUAUCUUCUUC-3′ (SEQ ID NO: 3063) AAT-496 19 nt Target #3: 5′-ACAGCACCAAUAUCUUCUU-3′ (SEQ ID NO: 3203) AAT-497 19 nt Target #1: 5′-GCACCAAUAUCUUCUUCUC-3′ (SEQ ID NO: 2924) AAT-497 19 nt Target #2: 5′-AGCACCAAUAUCUUCUUCU-3′ (SEQ ID NO: 3064) AAT-497 19 nt Target #3: 5′-CAGCACCAAUAUCUUCUUC-3′ (SEQ ID NO: 3204) AAT-498 19 nt Target #1: 5′-CACCAAUAUCUUCUUCUCC-3′ (SEQ ID NO: 2925) AAT-498 19 nt Target #2: 5′-GCACCAAUAUCUUCUUCUC-3′ (SEQ ID NO: 3065) AAT-498 19 nt Target #3: 5′-AGCACCAAUAUCUUCUUCU-3′ (SEQ ID NO: 3205) AAT-499 19 nt Target #1: 5′-ACCAAUAUCUUCUUCUCCC-3′ (SEQ ID NO: 2926) AAT-499 19 nt Target #2: 5′-CACCAAUAUCUUCUUCUCC-3′ (SEQ ID NO: 3066) AAT-499 19 nt Target #3: 5′-GCACCAAUAUCUUCUUCUC-3′ (SEQ ID NO: 3206) AAT-516 19 nt Target #1: 5′-CCCAGUGAGCAUCGCUACA-3′ (SEQ ID NO: 2927) AAT-516 19 nt Target #2: 5′-CCCCAGUGAGCAUCGCUAC-3′ (SEQ ID NO: 3067) AAT-516 19 nt Target #3: 5′-UCCCCAGUGAGCAUCGCUA-3′ (SEQ ID NO: 3207) AAT-517 19 nt Target #1: 5′-CCAGUGAGCAUCGCUACAG-3′ (SEQ ID NO: 2928) AAT-517 19 nt Target #2: 5′-CCCAGUGAGCAUCGCUACA-3′ (SEQ ID NO: 3068) AAT-517 19 nt Target #3: 5′-CCCCAGUGAGCAUCGCUAC-3′ (SEQ ID NO: 3208) AAT-518 19 nt Target #1: 5′-CAGUGAGCAUCGCUACAGC-3′ (SEQ ID NO: 2929) AAT-518 19 nt Target #2: 5′-CCAGUGAGCAUCGCUACAG-3′ (SEQ ID NO: 3069) AAT-518 19 nt Target #3: 5′-CCCAGUGAGCAUCGCUACA-3′ (SEQ ID NO: 3209) AAT-519 19 nt Target #1: 5′-AGUGAGCAUCGCUACAGCC-3′ (SEQ ID NO: 2930) AAT-519 19 nt Target #2: 5′-CAGUGAGCAUCGCUACAGC-3′ (SEQ ID NO: 3070) AAT-519 19 nt Target #3: 5′-CCAGUGAGCAUCGCUACAG-3′ (SEQ ID NO: 3210) AAT-520 19 nt Target #1: 5′-GUGAGCAUCGCUACAGCCU-3′ (SEQ ID NO: 2931) AAT-520 19 nt Target #2: 5′-AGUGAGCAUCGCUACAGCC-3′ (SEQ ID NO: 3071) AAT-520 19 nt Target #3: 5′-CAGUGAGCAUCGCUACAGC-3′ (SEQ ID NO: 3211) AAT-521 19 nt Target #1: 5′-UGAGCAUCGCUACAGCCUU-3′ (SEQ ID NO: 2932) AAT-521 19 nt Target #2: 5′-GUGAGCAUCGCUACAGCCU-3′ (SEQ ID NO: 3072) AAT-521 19 nt Target #3: 5′-AGUGAGCAUCGCUACAGCC-3′ (SEQ ID NO: 3212) AAT-522 19 nt Target #1: 5′-GAGCAUCGCUACAGCCUUU-3′ (SEQ ID NO: 2933) AAT-522 19 nt Target #2: 5′-UGAGCAUCGCUACAGCCUU-3′ (SEQ ID NO: 3073) AAT-522 19 nt Target #3: 5′-GUGAGCAUCGCUACAGCCU-3′ (SEQ ID NO: 3213) AAT-523 19 nt Target #1: 5′-AGCAUCGCUACAGCCUUUG-3′ (SEQ ID NO: 2934) AAT-523 19 nt Target #2: 5′-GAGCAUCGCUACAGCCUUU-3′ (SEQ ID NO: 3074) AAT-523 19 nt Target #3: 5′-UGAGCAUCGCUACAGCCUU-3′ (SEQ ID NO: 3214) AAT-524 19 nt Target #1: 5′-GCAUCGCUACAGCCUUUGC-3′ (SEQ ID NO: 2935) AAT-524 19 nt Target #2: 5′-AGCAUCGCUACAGCCUUUG-3′ (SEQ ID NO: 3075) AAT-524 19 nt Target #3: 5′-GAGCAUCGCUACAGCCUUU-3′ (SEQ ID NO: 3215) AAT-525 19 nt Target #1: 5′-CAUCGCUACAGCCUUUGCA-3′ (SEQ ID NO: 2936) AAT-525 19 nt Target #2: 5′-GCAUCGCUACAGCCUUUGC-3′ (SEQ ID NO: 3076) AAT-525 19 nt Target #3: 5′-AGCAUCGCUACAGCCUUUG-3′ (SEQ ID NO: 3216) AAT-526 19 nt Target #1: 5′-AUCGCUACAGCCUUUGCAA-3′ (SEQ ID NO: 2937) AAT-526 19 nt Target #2: 5′-CAUCGCUACAGCCUUUGCA-3′ (SEQ ID NO: 3077) AAT-526 19 nt Target #3: 5′-GCAUCGCUACAGCCUUUGC-3′ (SEQ ID NO: 3217) AAT-527 19 nt Target #1: 5′-UCGCUACAGCCUUUGCAAU-3′ (SEQ ID NO: 2938) AAT-527 19 nt Target #2: 5′-AUCGCUACAGCCUUUGCAA-3′ (SEQ ID NO: 3078) AAT-527 19 nt Target #3: 5′-CAUCGCUACAGCCUUUGCA-3′ (SEQ ID NO: 3218) AAT-528 19 nt Target #1: 5′-CGCUACAGCCUUUGCAAUG-3′ (SEQ ID NO: 2939) AAT-528 19 nt Target #2: 5′-UCGCUACAGCCUUUGCAAU-3′ (SEQ ID NO: 3079) AAT-528 19 nt Target #3: 5′-AUCGCUACAGCCUUUGCAA-3′ (SEQ ID NO: 3219) AAT-529 19 nt Target #1: 5′-GCUACAGCCUUUGCAAUGC-3′ (SEQ ID NO: 2940) AAT-529 19 nt Target #2: 5′-CGCUACAGCCUUUGCAAUG-3′ (SEQ ID NO: 3080) AAT-529 19 nt Target #3: 5′-UCGCUACAGCCUUUGCAAU-3′ (SEQ ID NO: 3220) AAT-530 19 nt Target #1: 5′-CUACAGCCUUUGCAAUGCU-3′ (SEQ ID NO: 2941) AAT-530 19 nt Target #2: 5′-GCUACAGCCUUUGCAAUGC-3′ (SEQ ID NO: 3081) AAT-530 19 nt Target #3: 5′-CGCUACAGCCUUUGCAAUG-3′ (SEQ ID NO: 3221) AAT-531 19 nt Target #1: 5′-UACAGCCUUUGCAAUGCUC-3′ (SEQ ID NO: 2942) AAT-531 19 nt Target #2: 5′-CUACAGCCUUUGCAAUGCU-3′ (SEQ ID NO: 3082) AAT-531 19 nt Target #3: 5′-GCUACAGCCUUUGCAAUGC-3′ (SEQ ID NO: 3222) AAT-552 19 nt Target #1: 5′-CCUGGGGACCAAGGCUGAC-3′ (SEQ ID NO: 2943) AAT-552 19 nt Target #2: 5′-CCCUGGGGACCAAGGCUGA-3′ (SEQ ID NO: 3083) AAT-552 19 nt Target #3: 5′-UCCCUGGGGACCAAGGCUG-3′ (SEQ ID NO: 3223) AAT-556 19 nt Target #1: 5′-GGGACCAAGGCUGACACUC-3′ (SEQ ID NO: 2944) AAT-556 19 nt Target #2: 5′-GGGGACCAAGGCUGACACU-3′ (SEQ ID NO: 3084) AAT-556 19 nt Target #3: 5′-UGGGGACCAAGGCUGACAC-3′ (SEQ ID NO: 3224) AAT-557 19 nt Target #1: 5′-GGACCAAGGCUGACACUCA-3′ (SEQ ID NO: 2945) AAT-557 19 nt Target #2: 5′-GGGACCAAGGCUGACACUC-3′ (SEQ ID NO: 3085) AAT-557 19 nt Target #3: 5′-GGGGACCAAGGCUGACACU-3′ (SEQ ID NO: 3225) AAT-558 19 nt Target #1: 5′-GACCAAGGCUGACACUCAC-3′ (SEQ ID NO: 2946) AAT-558 19 nt Target #2: 5′-GGACCAAGGCUGACACUCA-3′ (SEQ ID NO: 3086) AAT-558 19 nt Target #3: 5′-GGGACCAAGGCUGACACUC-3′ (SEQ ID NO: 3226) AAT-579 19 nt Target #1: 5′-UGAAAUCCUGGAGGGCCUG-3′ (SEQ ID NO: 2947) AAT-579 19 nt Target #2: 5′-AUGAAAUCCUGGAGGGCCU-3′ (SEQ ID NO: 3087) AAT-579 19 nt Target #3: 5′-GAUGAAAUCCUGGAGGGCC-3′ (SEQ ID NO: 3227) AAT-580 19 nt Target #1: 5′-GAAAUCCUGGAGGGCCUGA-3′ (SEQ ID NO: 2948) AAT-580 19 nt Target #2: 5′-UGAAAUCCUGGAGGGCCUG-3′ (SEQ ID NO: 3088) AAT-580 19 nt Target #3: 5′-AUGAAAUCCUGGAGGGCCU-3′ (SEQ ID NO: 3228) AAT-632 19 nt Target #1: 5′-UCCAUGAAGGCUUCCAGGA-3′ (SEQ ID NO: 2949) AAT-632 19 nt Target #2: 5′-AUCCAUGAAGGCUUCCAGG-3′ (SEQ ID NO: 3089) AAT-632 19 nt Target #3: 5′-GAUCCAUGAAGGCUUCCAG-3′ (SEQ ID NO: 3229) AAT-633 19 nt Target #1: 5′-CCAUGAAGGCUUCCAGGAA-3′ (SEQ ID NO: 2950) AAT-633 19 nt Target #2: 5′-UCCAUGAAGGCUUCCAGGA-3′ (SEQ ID NO: 3090) AAT-633 19 nt Target #3: 5′-AUCCAUGAAGGCUUCCAGG-3′ (SEQ ID NO: 3230) AAT-801 19 nt Target #1: 5′-GGACACCGAAGAGGCCAAG-3′ (SEQ ID NO: 2951) AAT-801 19 nt Target #2: 5′-GGGACACCGAAGAGGCCAA-3′ (SEQ ID NO: 3091) AAT-801 19 nt Target #3: 5′-GGGGACACCGAAGAGGCCA-3′ (SEQ ID NO: 3231) AAT-802 19 nt Target #1: 5′-GACACCGAAGAGGCCAAGA-3′ (SEQ ID NO: 2952) AAT-802 19 nt Target #2: 5′-GGACACCGAAGAGGCCAAG-3′ (SEQ ID NO: 3092) AAT-802 19 nt Target #3: 5′-GGGACACCGAAGAGGCCAA-3′ (SEQ ID NO: 3232) AAT-803 19 nt Target #1: 5′-ACACCGAAGAGGCCAAGAA-3′ (SEQ ID NO: 2953) AAT-803 19 nt Target #2: 5′-GACACCGAAGAGGCCAAGA-3′ (SEQ ID NO: 3093) AAT-803 19 nt Target #3: 5′-GGACACCGAAGAGGCCAAG-3′ (SEQ ID NO: 3233) AAT-804 19 nt Target #1: 5′-CACCGAAGAGGCCAAGAAA-3′ (SEQ ID NO: 2954) AAT-804 19 nt Target #2: 5′-ACACCGAAGAGGCCAAGAA-3′ (SEQ ID NO: 3094) AAT-804 19 nt Target #3: 5′-GACACCGAAGAGGCCAAGA-3′ (SEQ ID NO: 3234) AAT-805 19 nt Target #1: 5′-ACCGAAGAGGCCAAGAAAC-3′ (SEQ ID NO: 2955) AAT-805 19 nt Target #2: 5′-CACCGAAGAGGCCAAGAAA-3′ (SEQ ID NO: 3095) AAT-805 19 nt Target #3: 5′-ACACCGAAGAGGCCAAGAA-3′ (SEQ ID NO: 3235) AAT-806 19 nt Target #1: 5′-CCGAAGAGGCCAAGAAACA-3′ (SEQ ID NO: 2956) AAT-806 19 nt Target #2: 5′-ACCGAAGAGGCCAAGAAAC-3′ (SEQ ID NO: 3096) AAT-806 19 nt Target #3: 5′-CACCGAAGAGGCCAAGAAA-3′ (SEQ ID NO: 3236) AAT-807 19 nt Target #1: 5′-CGAAGAGGCCAAGAAACAG-3′ (SEQ ID NO: 2957) AAT-807 19 nt Target #2: 5′-CCGAAGAGGCCAAGAAACA-3′ (SEQ ID NO: 3097) AAT-807 19 nt Target #3: 5′-ACCGAAGAGGCCAAGAAAC-3′ (SEQ ID NO: 3237) AAT-808 19 nt Target #1: 5′-GAAGAGGCCAAGAAACAGA-3′ (SEQ ID NO: 2958) AAT-808 19 nt Target #2: 5′-CGAAGAGGCCAAGAAACAG-3′ (SEQ ID NO: 3098) AAT-808 19 nt Target #3: 5′-CCGAAGAGGCCAAGAAACA-3′ (SEQ ID NO: 3238) AAT-809 19 nt Target #1: 5′-AAGAGGCCAAGAAACAGAU-3′ (SEQ ID NO: 2959) AAT-809 19 nt Target #2: 5′-GAAGAGGCCAAGAAACAGA-3′ (SEQ ID NO: 3099) AAT-809 19 nt Target #3: 5′-CGAAGAGGCCAAGAAACAG-3′ (SEQ ID NO: 3239) AAT-810 19 nt Target #1: 5′-AGAGGCCAAGAAACAGAUC-3′ (SEQ ID NO: 2960) AAT-810 19 nt Target #2: 5′-AAGAGGCCAAGAAACAGAU-3′ (SEQ ID NO: 3100) AAT-810 19 nt Target #3: 5′-GAAGAGGCCAAGAAACAGA-3′ (SEQ ID NO: 3240) AAT-811 19 nt Target #1: 5′-GAGGCCAAGAAACAGAUCA-3′ (SEQ ID NO: 2961) AAT-811 19 nt Target #2: 5′-AGAGGCCAAGAAACAGAUC-3′ (SEQ ID NO: 3101) AAT-811 19 nt Target #3: 5′-AAGAGGCCAAGAAACAGAU-3′ (SEQ ID NO: 3241) AAT-812 19 nt Target #1: 5′-AGGCCAAGAAACAGAUCAA-3′ (SEQ ID NO: 2962) AAT-812 19 nt Target #2: 5′-GAGGCCAAGAAACAGAUCA-3′ (SEQ ID NO: 3102) AAT-812 19 nt Target #3: 5′-AGAGGCCAAGAAACAGAUC-3′ (SEQ ID NO: 3242) AAT-813 19 nt Target #1: 5′-GGCCAAGAAACAGAUCAAC-3′ (SEQ ID NO: 2963) AAT-813 19 nt Target #2: 5′-AGGCCAAGAAACAGAUCAA-3′ (SEQ ID NO: 3103) AAT-813 19 nt Target #3: 5′-GAGGCCAAGAAACAGAUCA-3′ (SEQ ID NO: 3243) AAT-900 19 nt Target #1: 5′-UUUUGCUCUGGUGAAUUAC-3′ (SEQ ID NO: 2964) AAT-900 19 nt Target #2: 5′-UUUUUGCUCUGGUGAAUUA-3′ (SEQ ID NO: 3104) AAT-900 19 nt Target #3: 5′-GUUUUUGCUCUGGUGAAUU-3′ (SEQ ID NO: 3244) AAT-901 19 nt Target #1: 5′-UUUGCUCUGGUGAAUUACA-3′ (SEQ ID NO: 2965) AAT-901 19 nt Target #2: 5′-UUUUGCUCUGGUGAAUUAC-3′ (SEQ ID NO: 3105) AAT-901 19 nt Target #3: 5′-UUUUUGCUCUGGUGAAUUA-3′ (SEQ ID NO: 3245) AAT-902 19 nt Target #1: 5′-UUGCUCUGGUGAAUUACAU-3′ (SEQ ID NO: 2966) AAT-902 19 nt Target #2: 5′-UUUGCUCUGGUGAAUUACA-3′ (SEQ ID NO: 3106) AAT-902 19 nt Target #3: 5′-UUUUGCUCUGGUGAAUUAC-3′ (SEQ ID NO: 3246) AAT-903 19 nt Target #1: 5′-UGCUCUGGUGAAUUACAUC-3′ (SEQ ID NO: 2967) AAT-903 19 nt Target #2: 5′-UUGCUCUGGUGAAUUACAU-3′ (SEQ ID NO: 3107) AAT-903 19 nt Target #3: 5′-UUUGCUCUGGUGAAUUACA-3′ (SEQ ID NO: 3247) AAT-904 19 nt Target #1: 5′-GCUCUGGUGAAUUACAUCU-3′ (SEQ ID NO: 2968) AAT-904 19 nt Target #2: 5′-UGCUCUGGUGAAUUACAUC-3′ (SEQ ID NO: 3108) AAT-904 19 nt Target #3: 5′-UUGCUCUGGUGAAUUACAU-3′ (SEQ ID NO: 3248) AAT-905 19 nt Target #1: 5′-CUCUGGUGAAUUACAUCUU-3′ (SEQ ID NO: 2969) AAT-905 19 nt Target #2: 5′-GCUCUGGUGAAUUACAUCU-3′ (SEQ ID NO: 3109) AAT-905 19 nt Target #3: 5′-UGCUCUGGUGAAUUACAUC-3′ (SEQ ID NO: 3249) AAT-906 19 nt Target #1: 5′-UCUGGUGAAUUACAUCUUC-3′ (SEQ ID NO: 2970) AAT-906 19 nt Target #2: 5′-CUCUGGUGAAUUACAUCUU-3′ (SEQ ID NO: 3110) AAT-906 19 nt Target #3: 5′-GCUCUGGUGAAUUACAUCU-3′ (SEQ ID NO: 3250) AAT-907 19 nt Target #1: 5′-CUGGUGAAUUACAUCUUCU-3′ (SEQ ID NO: 2971) AAT-907 19 nt Target #2: 5′-UCUGGUGAAUUACAUCUUC-3′ (SEQ ID NO: 3111) AAT-907 19 nt Target #3: 5′-CUCUGGUGAAUUACAUCUU-3′ (SEQ ID NO: 3251) AAT-908 19 nt Target #1: 5′-UGGUGAAUUACAUCUUCUU-3′ (SEQ ID NO: 2972) AAT-908 19 nt Target #2: 5′-CUGGUGAAUUACAUCUUCU-3′ (SEQ ID NO: 3112) AAT-908 19 nt Target #3: 5′-UCUGGUGAAUUACAUCUUC-3′ (SEQ ID NO: 3252) AAT-909 19 nt Target #1: 5′-GGUGAAUUACAUCUUCUUU-3′ (SEQ ID NO: 2973) AAT-909 19 nt Target #2: 5′-UGGUGAAUUACAUCUUCUU-3′ (SEQ ID NO: 3113) AAT-909 19 nt Target #3: 5′-CUGGUGAAUUACAUCUUCU-3′ (SEQ ID NO: 3253) AAT-910 19 nt Target #1: 5′-GUGAAUUACAUCUUCUUUA-3′ (SEQ ID NO: 2974) AAT-910 19 nt Target #2: 5′-GGUGAAUUACAUCUUCUUU-3′ (SEQ ID NO: 3114) AAT-910 19 nt Target #3: 5′-UGGUGAAUUACAUCUUCUU-3′ (SEQ ID NO: 3254) AAT-911 19 nt Target #1: 5′-UGAAUUACAUCUUCUUUAA-3′ (SEQ ID NO: 2975) AAT-911 19 nt Target #2: 5′-GUGAAUUACAUCUUCUUUA-3′ (SEQ ID NO: 3115) AAT-911 19 nt Target #3: 5′-GGUGAAUUACAUCUUCUUU-3′ (SEQ ID NO: 3255) AAT-912 19 nt Target #1: 5′-GAAUUACAUCUUCUUUAAA-3′ (SEQ ID NO: 2976) AAT-912 19 nt Target #2: 5′-UGAAUUACAUCUUCUUUAA-3′ (SEQ ID NO: 3116) AAT-912 19 nt Target #3: 5′-GUGAAUUACAUCUUCUUUA-3′ (SEQ ID NO: 3256) AAT-913 19 nt Target #1: 5′-AAUUACAUCUUCUUUAAAG-3′ (SEQ ID NO: 2977) AAT-913 19 nt Target #2: 5′-GAAUUACAUCUUCUUUAAA-3′ (SEQ ID NO: 3117) AAT-913 19 nt Target #3: 5′-UGAAUUACAUCUUCUUUAA-3′ (SEQ ID NO: 3257) AAT-914 19 nt Target #1: 5′-AUUACAUCUUCUUUAAAGG-3′ (SEQ ID NO: 2978) AAT-914 19 nt Target #2: 5′-AAUUACAUCUUCUUUAAAG-3′ (SEQ ID NO: 3118) AAT-914 19 nt Target #3: 5′-GAAUUACAUCUUCUUUAAA-3′ (SEQ ID NO: 3258) AAT-915 19 nt Target #1: 5′-UUACAUCUUCUUUAAAGGC-3′ (SEQ ID NO: 2979) AAT-915 19 nt Target #2: 5′-AUUACAUCUUCUUUAAAGG-3′ (SEQ ID NO: 3119) AAT-915 19 nt Target #3: 5′-AAUUACAUCUUCUUUAAAG-3′ (SEQ ID NO: 3259) AAT-916 19 nt Target #1: 5′-UACAUCUUCUUUAAAGGCA-3′ (SEQ ID NO: 2980) AAT-916 19 nt Target #2: 5′-UUACAUCUUCUUUAAAGGC-3′ (SEQ ID NO: 3120) AAT-916 19 nt Target #3: 5′-AUUACAUCUUCUUUAAAGG-3′ (SEQ ID NO: 3260) AAT-917 19 nt Target #1: 5′-ACAUCUUCUUUAAAGGCAA-3′ (SEQ ID NO: 2981) AAT-917 19 nt Target #2: 5′-UACAUCUUCUUUAAAGGCA-3′ (SEQ ID NO: 3121) AAT-917 19 nt Target #3: 5′-UUACAUCUUCUUUAAAGGC-3′ (SEQ ID NO: 3261) AAT-918 19 nt Target #1: 5′-CAUCUUCUUUAAAGGCAAA-3′ (SEQ ID NO: 2982) AAT-918 19 nt Target #2: 5′-ACAUCUUCUUUAAAGGCAA-3′ (SEQ ID NO: 3122) AAT-918 19 nt Target #3: 5′-UACAUCUUCUUUAAAGGCA-3′ (SEQ ID NO: 3262) AAT-922 19 nt Target #1: 5′-UUCUUUAAAGGCAAAUGGG-3′ (SEQ ID NO: 2983) AAT-922 19 nt Target #2: 5′-CUUCUUUAAAGGCAAAUGG-3′ (SEQ ID NO: 3123) AAT-922 19 nt Target #3: 5′-UCUUCUUUAAAGGCAAAUG-3′ (SEQ ID NO: 3263) AAT-924 19 nt Target #1: 5′-CUUUAAAGGCAAAUGGGAG-3′ (SEQ ID NO: 2984) AAT-924 19 nt Target #2: 5′-UCUUUAAAGGCAAAUGGGA-3′ (SEQ ID NO: 3124) AAT-924 19 nt Target #3: 5′-UUCUUUAAAGGCAAAUGGG-3′ (SEQ ID NO: 3264) AAT-932 19 nt Target #1: 5′-GCAAAUGGGAGAGACCCUU-3′ (SEQ ID NO: 2985) AAT-932 19 nt Target #2: 5′-GGCAAAUGGGAGAGACCCU-3′ (SEQ ID NO: 3125) AAT-932 19 nt Target #3: 5′-AGGCAAAUGGGAGAGACCC-3′ (SEQ ID NO: 3265) AAT-933 19 nt Target #1: 5′-CAAAUGGGAGAGACCCUUU-3′ (SEQ ID NO: 2986) AAT-933 19 nt Target #2: 5′-GCAAAUGGGAGAGACCCUU-3′ (SEQ ID NO: 3126) AAT-933 19 nt Target #3: 5′-GGCAAAUGGGAGAGACCCU-3′ (SEQ ID NO: 3266) AAT-934 19 nt Target #1: 5′-AAAUGGGAGAGACCCUUUG-3′ (SEQ ID NO: 2987) AAT-934 19 nt Target #2: 5′-CAAAUGGGAGAGACCCUUU-3′ (SEQ ID NO: 3127) AAT-934 19 nt Target #3: 5′-GCAAAUGGGAGAGACCCUU-3′ (SEQ ID NO: 3267) AAT-935 19 nt Target #1: 5′-AAUGGGAGAGACCCUUUGA-3′ (SEQ ID NO: 2988) AAT-935 19 nt Target #2: 5′-AAAUGGGAGAGACCCUUUG-3′ (SEQ ID NO: 3128) AAT-935 19 nt Target #3: 5′-CAAAUGGGAGAGACCCUUU-3′ (SEQ ID NO: 3268) AAT-1061 19 nt Target #1: 5′-UGUCCAGCUGGGUGCUGCU-3′ (SEQ ID NO: 2989) AAT-1061 19 nt Target #2: 5′-CUGUCCAGCUGGGUGCUGC-3′ (SEQ ID NO: 3129) AAT-1061 19 nt Target #3: 5′-GCUGUCCAGCUGGGUGCUG-3′ (SEQ ID NO: 3269) AAT-1062 19 nt Target #1: 5′-GUCCAGCUGGGUGCUGCUG-3′ (SEQ ID NO: 2990) AAT-1062 19 nt Target #2: 5′-UGUCCAGCUGGGUGCUGCU-3′ (SEQ ID NO: 3130) AAT-1062 19 nt Target #3: 5′-CUGUCCAGCUGGGUGCUGC-3′ (SEQ ID NO: 3270) AAT-1063 19 nt Target #1: 5′-UCCAGCUGGGUGCUGCUGA-3′ (SEQ ID NO: 2991) AAT-1063 19 nt Target #2: 5′-GUCCAGCUGGGUGCUGCUG-3′ (SEQ ID NO: 3131) AAT-1063 19 nt Target #3: 5′-UGUCCAGCUGGGUGCUGCU-3′ (SEQ ID NO: 3271) AAT-1064 19 nt Target #1: 5′-CCAGCUGGGUGCUGCUGAU-3′ (SEQ ID NO: 2992) AAT-1064 19 nt Target #2: 5′-UCCAGCUGGGUGCUGCUGA-3′ (SEQ ID NO: 3132) AAT-1064 19 nt Target #3: 5′-GUCCAGCUGGGUGCUGCUG-3′ (SEQ ID NO: 3272) AAT-1065 19 nt Target #1: 5′-CAGCUGGGUGCUGCUGAUG-3′ (SEQ ID NO: 2993) AAT-1065 19 nt Target #2: 5′-CCAGCUGGGUGCUGCUGAU-3′ (SEQ ID NO: 3133) AAT-1065 19 nt Target #3: 5′-UCCAGCUGGGUGCUGCUGA-3′ (SEQ ID NO: 3273) AAT-1066 19 nt Target #1: 5′-AGCUGGGUGCUGCUGAUGA-3′ (SEQ ID NO: 2994) AAT-1066 19 nt Target #2: 5′-CAGCUGGGUGCUGCUGAUG-3′ (SEQ ID NO: 3134) AAT-1066 19 nt Target #3: 5′-CCAGCUGGGUGCUGCUGAU-3′ (SEQ ID NO: 3274) AAT-1067 19 nt Target #1: 5′-GCUGGGUGCUGCUGAUGAA-3′ (SEQ ID NO: 2995) AAT-1067 19 nt Target #2: 5′-AGCUGGGUGCUGCUGAUGA-3′ (SEQ ID NO: 3135) AAT-1067 19 nt Target #3: 5′-CAGCUGGGUGCUGCUGAUG-3′ (SEQ ID NO: 3275) AAT-1068 19 nt Target #1: 5′-CUGGGUGCUGCUGAUGAAA-3′ (SEQ ID NO: 2996) AAT-1068 19 nt Target #2: 5′-GCUGGGUGCUGCUGAUGAA-3′ (SEQ ID NO: 3136) AAT-1068 19 nt Target #3: 5′-AGCUGGGUGCUGCUGAUGA-3′ (SEQ ID NO: 3276) AAT-1069 19 nt Target #1: 5′-UGGGUGCUGCUGAUGAAAU-3′ (SEQ ID NO: 2997) AAT-1069 19 nt Target #2: 5′-CUGGGUGCUGCUGAUGAAA-3′ (SEQ ID NO: 3137) AAT-1069 19 nt Target #3: 5′-GCUGGGUGCUGCUGAUGAA-3′ (SEQ ID NO: 3277) AAT-1070 19 nt Target #1: 5′-GGGUGCUGCUGAUGAAAUA-3′ (SEQ ID NO: 2998) AAT-1070 19 nt Target #2: 5′-UGGGUGCUGCUGAUGAAAU-3′ (SEQ ID NO: 3138) AAT-1070 19 nt Target #3: 5′-CUGGGUGCUGCUGAUGAAA-3′ (SEQ ID NO: 3278) AAT-1072 19 nt Target #1: 5′-GUGCUGCUGAUGAAAUACC-3′ (SEQ ID NO: 2999) AAT-1072 19 nt Target #2: 5′-GGUGCUGCUGAUGAAAUAC-3′ (SEQ ID NO: 3139) AAT-1072 19 nt Target #3: 5′-GGGUGCUGCUGAUGAAAUA-3′ (SEQ ID NO: 3279) AAT-1073 19 nt Target #1: 5′-UGCUGCUGAUGAAAUACCU-3′ (SEQ ID NO: 3000) AAT-1073 19 nt Target #2: 5′-GUGCUGCUGAUGAAAUACC-3′ (SEQ ID NO: 3140) AAT-1073 19 nt Target #3: 5′-GGUGCUGCUGAUGAAAUAC-3′ (SEQ ID NO: 3280) AAT-1074 19 nt Target #1: 5′-GCUGCUGAUGAAAUACCUG-3′ (SEQ ID NO: 3001) AAT-1074 19 nt Target #2: 5′-UGCUGCUGAUGAAAUACCU-3′ (SEQ ID NO: 3141) AAT-1074 19 nt Target #3: 5′-GUGCUGCUGAUGAAAUACC-3′ (SEQ ID NO: 3281) AAT-1075 19 nt Target #1: 5′-CUGCUGAUGAAAUACCUGG-3′ (SEQ ID NO: 3002) AAT-1075 19 nt Target #2: 5′-GCUGCUGAUGAAAUACCUG-3′ (SEQ ID NO: 3142) AAT-1075 19 nt Target #3: 5′-UGCUGCUGAUGAAAUACCU-3′ (SEQ ID NO: 3282) AAT-1076 19 nt Target #1: 5′-UGCUGAUGAAAUACCUGGG-3′ (SEQ ID NO: 3003) AAT-1076 19 nt Target #2: 5′-CUGCUGAUGAAAUACCUGG-3′ (SEQ ID NO: 3143) AAT-1076 19 nt Target #3: 5′-GCUGCUGAUGAAAUACCUG-3′ (SEQ ID NO: 3283) AAT-1077 19 nt Target #1: 5′-GCUGAUGAAAUACCUGGGC-3′ (SEQ ID NO: 3004) AAT-1077 19 nt Target #2: 5′-UGCUGAUGAAAUACCUGGG-3′ (SEQ ID NO: 3144) AAT-1077 19 nt Target #3: 5′-CUGCUGAUGAAAUACCUGG-3′ (SEQ ID NO: 3284) AAT-1078 19 nt Target #1: 5′-CUGAUGAAAUACCUGGGCA-3′ (SEQ ID NO: 3005) AAT-1078 19 nt Target #2: 5′-GCUGAUGAAAUACCUGGGC-3′ (SEQ ID NO: 3145) AAT-1078 19 nt Target #3: 5′-UGCUGAUGAAAUACCUGGG-3′ (SEQ ID NO: 3285) AAT-1079 19 nt Target #1: 5′-UGAUGAAAUACCUGGGCAA-3′ (SEQ ID NO: 3006) AAT-1079 19 nt Target #2: 5′-CUGAUGAAAUACCUGGGCA-3′ (SEQ ID NO: 3146) AAT-1079 19 nt Target #3: 5′-GCUGAUGAAAUACCUGGGC-3′ (SEQ ID NO: 3286) AAT-1080 19 nt Target #1: 5′-GAUGAAAUACCUGGGCAAU-3′ (SEQ ID NO: 3007) AAT-1080 19 nt Target #2: 5′-UGAUGAAAUACCUGGGCAA-3′ (SEQ ID NO: 3147) AAT-1080 19 nt Target #3: 5′-CUGAUGAAAUACCUGGGCA-3′ (SEQ ID NO: 3287) AAT-1081 19 nt Target #1: 5′-AUGAAAUACCUGGGCAAUG-3′ (SEQ ID NO: 3008) AAT-1081 19 nt Target #2: 5′-GAUGAAAUACCUGGGCAAU-3′ (SEQ ID NO: 3148) AAT-1081 19 nt Target #3: 5′-UGAUGAAAUACCUGGGCAA-3′ (SEQ ID NO: 3288) AAT-1083 19 nt Target #1: 5′-GAAAUACCUGGGCAAUGCC-3′ (SEQ ID NO: 3009) AAT-1083 19 nt Target #2: 5′-UGAAAUACCUGGGCAAUGC-3′ (SEQ ID NO: 3149) AAT-1083 19 nt Target #3: 5′-AUGAAAUACCUGGGCAAUG-3′ (SEQ ID NO: 3289) AAT-1138 19 nt Target #1: 5′-CAGCACCUGGAAAAUGAAC-3′ (SEQ ID NO: 3010) AAT-1138 19 nt Target #2: 5′-ACAGCACCUGGAAAAUGAA-3′ (SEQ ID NO: 3150) AAT-1138 19 nt Target #3: 5′-UACAGCACCUGGAAAAUGA-3′ (SEQ ID NO: 3290) AAT-1144 19 nt Target #1: 5′-CUGGAAAAUGAACUCACCC-3′ (SEQ ID NO: 3011) AAT-1144 19 nt Target #2: 5′-CCUGGAAAAUGAACUCACC-3′ (SEQ ID NO: 3151) AAT-1144 19 nt Target #3: 5′-ACCUGGAAAAUGAACUCAC-3′ (SEQ ID NO: 3291) AAT-1145 19 nt Target #1: 5′-UGGAAAAUGAACUCACCCA-3′ (SEQ ID NO: 3012) AAT-1145 19 nt Target #2: 5′-CUGGAAAAUGAACUCACCC-3′ (SEQ ID NO: 3152) AAT-1145 19 nt Target #3: 5′-CCUGGAAAAUGAACUCACC-3′ (SEQ ID NO: 3292) AAT-1165 19 nt Target #1: 5′-GAUAUCAUCACCAAGUUCC-3′ (SEQ ID NO: 3013) AAT-1165 19 nt Target #2: 5′-CGAUAUCAUCACCAAGUUC-3′ (SEQ ID NO: 3153) AAT-1165 19 nt Target #3: 5′-ACGAUAUCAUCACCAAGUU-3′ (SEQ ID NO: 3293) AAT-1176 19 nt Target #1: 5′-CAAGUUCCUGGAAAAUGAA-3′ (SEQ ID NO: 3014) AAT-1176 19 nt Target #2: 5′-CCAAGUUCCUGGAAAAUGA-3′ (SEQ ID NO: 3154) AAT-1176 19 nt Target #3: 5′-ACCAAGUUCCUGGAAAAUG-3′ (SEQ ID NO: 3294) AAT-1232 19 nt Target #1: 5′-CCAUUACUGGAACCUAUGA-3′ (SEQ ID NO: 3015) AAT-1232 19 nt Target #2: 5′-UCCAUUACUGGAACCUAUG-3′ (SEQ ID NO: 3155) AAT-1232 19 nt Target #3: 5′-GUCCAUUACUGGAACCUAU-3′ (SEQ ID NO: 3295) AAT-1233 19 nt Target #1: 5′-CAUUACUGGAACCUAUGAU-3′ (SEQ ID NO: 3016) AAT-1233 19 nt Target #2: 5′-CCAUUACUGGAACCUAUGA-3′ (SEQ ID NO: 3156) AAT-1233 19 nt Target #3: 5′-UCCAUUACUGGAACCUAUG-3′ (SEQ ID NO: 3296) AAT-1234 19 nt Target #1: 5′-AUUACUGGAACCUAUGAUC-3′ (SEQ ID NO: 3017) AAT-1234 19 nt Target #2: 5′-CAUUACUGGAACCUAUGAU-3′ (SEQ ID NO: 3157) AAT-1234 19 nt Target #3: 5′-CCAUUACUGGAACCUAUGA-3′ (SEQ ID NO: 3297) AAT-1235 19 nt Target #1: 5′-UUACUGGAACCUAUGAUCU-3′ (SEQ ID NO: 3018) AAT-1235 19 nt Target #2: 5′-AUUACUGGAACCUAUGAUC-3′ (SEQ ID NO: 3158) AAT-1235 19 nt Target #3: 5′-CAUUACUGGAACCUAUGAU-3′ (SEQ ID NO: 3298) AAT-1236 19 nt Target #1: 5′-UACUGGAACCUAUGAUCUG-3′ (SEQ ID NO: 3019) AAT-1236 19 nt Target #2: 5′-UUACUGGAACCUAUGAUCU-3′ (SEQ ID NO: 3159) AAT-1236 19 nt Target #3: 5′-AUUACUGGAACCUAUGAUC-3′ (SEQ ID NO: 3299) AAT-1237 19 nt Target #1: 5′-ACUGGAACCUAUGAUCUGA-3′ (SEQ ID NO: 3020) AAT-1237 19 nt Target #2: 5′-UACUGGAACCUAUGAUCUG-3′ (SEQ ID NO: 3160) AAT-1237 19 nt Target #3: 5′-UUACUGGAACCUAUGAUCU-3′ (SEQ ID NO: 3300) AAT-1238 19 nt Target #1: 5′-CUGGAACCUAUGAUCUGAA-3′ (SEQ ID NO: 3021) AAT-1238 19 nt Target #2: 5′-ACUGGAACCUAUGAUCUGA-3′ (SEQ ID NO: 3161) AAT-1238 19 nt Target #3: 5′-UACUGGAACCUAUGAUCUG-3′ (SEQ ID NO: 3301) AAT-1239 19 nt Target #1: 5′-UGGAACCUAUGAUCUGAAG-3′ (SEQ ID NO: 3022) AAT-1239 19 nt Target #2: 5′-CUGGAACCUAUGAUCUGAA-3′ (SEQ ID NO: 3162) AAT-1239 19 nt Target #3: 5′-ACUGGAACCUAUGAUCUGA-3′ (SEQ ID NO: 3302) AAT-1240 19 nt Target #1: 5′-GGAACCUAUGAUCUGAAGA-3′ (SEQ ID NO: 3023) AAT-1240 19 nt Target #2: 5′-UGGAACCUAUGAUCUGAAG-3′ (SEQ ID NO: 3163) AAT-1240 19 nt Target #3: 5′-CUGGAACCUAUGAUCUGAA-3′ (SEQ ID NO: 3303) AAT-1279 19 nt Target #1: 5′-AUCACUAAGGUCUUCAGCA-3′ (SEQ ID NO: 3024) AAT-1279 19 nt Target #2: 5′-CAUCACUAAGGUCUUCAGC-3′ (SEQ ID NO: 3164) AAT-1279 19 nt Target #3: 5′-GCAUCACUAAGGUCUUCAG-3′ (SEQ ID NO: 3304) AAT-1280 19 nt Target #1: 5′-UCACUAAGGUCUUCAGCAA-3′ (SEQ ID NO: 3025) AAT-1280 19 nt Target #2: 5′-AUCACUAAGGUCUUCAGCA-3′ (SEQ ID NO: 3165) AAT-1280 19 nt Target #3: 5′-CAUCACUAAGGUCUUCAGC-3′ (SEQ ID NO: 3305) AAT-1281 19 nt Target #1: 5′-CACUAAGGUCUUCAGCAAU-3′ (SEQ ID NO: 3026) AAT-1281 19 nt Target #2: 5′-UCACUAAGGUCUUCAGCAA-3′ (SEQ ID NO: 3166) AAT-1281 19 nt Target #3: 5′-AUCACUAAGGUCUUCAGCA-3′ (SEQ ID NO: 3306) AAT-1283 19 nt Target #1: 5′-CUAAGGUCUUCAGCAAUGG-3′ (SEQ ID NO: 3027) AAT-1283 19 nt Target #2: 5′-ACUAAGGUCUUCAGCAAUG-3′ (SEQ ID NO: 3167) AAT-1283 19 nt Target #3: 5′-CACUAAGGUCUUCAGCAAU-3′ (SEQ ID NO: 3307) AAT-1284 19 nt Target #1: 5′-UAAGGUCUUCAGCAAUGGG-3′ (SEQ ID NO: 3028) AAT-1284 19 nt Target #2: 5′-CUAAGGUCUUCAGCAAUGG-3′ (SEQ ID NO: 3168) AAT-1284 19 nt Target #3: 5′-ACUAAGGUCUUCAGCAAUG-3′ (SEQ ID NO: 3308) AAT-1337 19 nt Target #1: 5′-UGAAGCUCUCCAAGGCCGU-3′ (SEQ ID NO: 3029) AAT-1337 19 nt Target #2: 5′-CUGAAGCUCUCCAAGGCCG-3′ (SEQ ID NO: 3169) AAT-1337 19 nt Target #3: 5′-CCUGAAGCUCUCCAAGGCC-3′ (SEQ ID NO: 3309) AAT-1338 19 nt Target #1: 5′-GAAGCUCUCCAAGGCCGUG-3′ (SEQ ID NO: 3030) AAT-1338 19 nt Target #2: 5′-UGAAGCUCUCCAAGGCCGU-3′ (SEQ ID NO: 3170) AAT-1338 19 nt Target #3: 5′-CUGAAGCUCUCCAAGGCCG-3′ (SEQ ID NO: 3310) AAT-1339 19 nt Target #1: 5′-AAGCUCUCCAAGGCCGUGC-3′ (SEQ ID NO: 3031) AAT-1339 19 nt Target #2: 5′-GAAGCUCUCCAAGGCCGUG-3′ (SEQ ID NO: 3171) AAT-1339 19 nt Target #3: 5′-UGAAGCUCUCCAAGGCCGU-3′ (SEQ ID NO: 3311) AAT-1442 19 nt Target #1: 5′-CCGAGGUCAAGUUCAACAA-3′ (SEQ ID NO: 3032) AAT-1442 19 nt Target #2: 5′-CCCGAGGUCAAGUUCAACA-3′ (SEQ ID NO: 3172) AAT-1442 19 nt Target #3: 5′-CCCCGAGGUCAAGUUCAAC-3′ (SEQ ID NO: 3312) AAT-1443 19 nt Target #1: 5′-CGAGGUCAAGUUCAACAAA-3′ (SEQ ID NO: 3033) AAT-1443 19 nt Target #2: 5′-CCGAGGUCAAGUUCAACAA-3′ (SEQ ID NO: 3173) AAT-1443 19 nt Target #3: 5′-CCCGAGGUCAAGUUCAACA-3′ (SEQ ID NO: 3313) AAT-1444 19 nt Target #1: 5′-GAGGUCAAGUUCAACAAAC-3′ (SEQ ID NO: 3034) AAT-1444 19 nt Target #2: 5′-CGAGGUCAAGUUCAACAAA-3′ (SEQ ID NO: 3174) AAT-1444 19 nt Target #3: 5′-CCGAGGUCAAGUUCAACAA-3′ (SEQ ID NO: 3314) AAT-1445 19 nt Target #1: 5′-AGGUCAAGUUCAACAAACC-3′ (SEQ ID NO: 3035) AAT-1445 19 nt Target #2: 5′-GAGGUCAAGUUCAACAAAC-3′ (SEQ ID NO: 3175) AAT-1445 19 nt Target #3: 5′-CGAGGUCAAGUUCAACAAA-3′ (SEQ ID NO: 3315) AAT-1446 19 nt Target #1: 5′-GGUCAAGUUCAACAAACCC-3′ (SEQ ID NO: 3036) AAT-1446 19 nt Target #2: 5′-AGGUCAAGUUCAACAAACC-3′ (SEQ ID NO: 3176) AAT-1446 19 nt Target #3: 5′-GAGGUCAAGUUCAACAAAC-3′ (SEQ ID NO: 3316) AAT-1447 19 nt Target #1: 5′-GUCAAGUUCAACAAACCCU-3′ (SEQ ID NO: 3037) AAT-1447 19 nt Target #2: 5′-GGUCAAGUUCAACAAACCC-3′ (SEQ ID NO: 3177) AAT-1447 19 nt Target #3: 5′-AGGUCAAGUUCAACAAACC-3′ (SEQ ID NO: 3317) AAT-1448 19 nt Target #1: 5′-UCAAGUUCAACAAACCCUU-3′ (SEQ ID NO: 3038) AAT-1448 19 nt Target #2: 5′-GUCAAGUUCAACAAACCCU-3′ (SEQ ID NO: 3178) AAT-1448 19 nt Target #3: 5′-GGUCAAGUUCAACAAACCC-3′ (SEQ ID NO: 3318) AAT-1449 19 nt Target #1: 5′-CAAGUUCAACAAACCCUUU-3′ (SEQ ID NO: 3039) AAT-1449 19 nt Target #2: 5′-UCAAGUUCAACAAACCCUU-3′ (SEQ ID NO: 3179) AAT-1449 19 nt Target #3: 5′-GUCAAGUUCAACAAACCCU-3′ (SEQ ID NO: 3319) AAT-1450 19 nt Target #1: 5′-AAGUUCAACAAACCCUUUG-3′ (SEQ ID NO: 3040) AAT-1450 19 nt Target #2: 5′-CAAGUUCAACAAACCCUUU-3′ (SEQ ID NO: 3180) AAT-1450 19 nt Target #3: 5′-UCAAGUUCAACAAACCCUU-3′ (SEQ ID NO: 3320) AAT-1451 19 nt Target #1: 5′-AGUUCAACAAACCCUUUGU-3′ (SEQ ID NO: 3041) AAT-1451 19 nt Target #2: 5′-AAGUUCAACAAACCCUUUG-3′ (SEQ ID NO: 3181) AAT-1451 19 nt Target #3: 5′-CAAGUUCAACAAACCCUUU-3′ (SEQ ID NO: 3321)

TABLE 17 Other Human Anti-+60 -1 antitrypsin DsiRNA Agents (Asymmetries) 5′-CCCCAGGGAGAUGCUGCCCAGAAga-3′ (SEQ ID NO: 3358) 3′-UAGGGGUCCCUCUACGACGGGUCUUCU-5′ (SEQ ID NO: 3373) AAT-364 Target: 5′-ATCCCCAGGGAGATGCTGCCCAGAAGA-3′ (SEQ ID NO: 3388) 5′-GCCUUUGCAAUGCUCUCCCUGGGga-3′ (SEQ ID NO: 3359) 3′-GUCGGAAACGUUACGAGAGGGACCCCU-5′ (SEQ ID NO: 3374) AAT-535 Target: 5′-CAGCCTTTGCAATGCTCTCCCTGGGGA-3′ (SEQ ID NO: 3389) 5′-GCAAUGCUCUCCCUGGGGACCAAgg-3′ (SEQ ID NO: 3360) 3′-AACGUUACGAGAGGGACCCCUGGUUCC-5′ (SEQ ID NO: 3375) AAT-541 Target: 5′-TTGCAATGCTCTCCCTGGGGACCAAGG-3′ (SEQ ID NO: 3390) 5′-GGGGACCAAGGCUGACACUCACGat-3′ (SEQ ID NO: 3361) 3′-GACCCCUGGUUCCGACUGUGAGUGCUA-5′ (SEQ ID NO: 3376) AAT-555 Target: 5′-CTGGGGACCAAGGCTGACACTCACGAT-3′ (SEQ ID NO: 3391) 5′-UCCUGGAGGGCCUGAAUUUCAACct-3′ (SEQ ID NO: 3362) 3′-UUAGGACCUCCCGGACUUAAAGUUGGA-5′ (SEQ ID NO: 3377) AAT-584 Target: 5′-AATCCTGGAGGGCCTGAATTTCAACCT-3′ (SEQ ID NO: 3392) 5′-CAGACAGCCAGCUCCAGCUGACCac-3′ (SEQ ID NO: 3363) 3′-CGGUCUGUCGGUCGAGGUCGACUGGUG-5′ (SEQ ID NO: 3378) AAT-674 Target: 5′-GCCAGACAGCCAGCTCCAGCTGACCAC-3′ (SEQ ID NO: 3393) 5′-AUCUUCUUUAAAGGCAAAUGGGAga-3′ (SEQ ID NO: 3364) 3′-UGUAGAAGAAAUUUCCGUUUACCCUCU-5′ (SEQ ID NO: 3379) AAT-919 Target: 5′-ACATCTTCTTTAAAGGCAAATGGGAGA-3′ (SEQ ID NO: 3394) 5′-UUAAAGGCAAAUGGGAGAGACCCtt-3′ (SEQ ID NO: 3365) 3′-GAAAUUUCCGUUUACCCUCUCUGGGAA-5′ (SEQ ID NO: 3380) AAT-926 Target: 5′-CTTTAAAGGCAAATGGGAGAGACCCTT-3′ (SEQ ID NO: 3395) 5′-UAAAGGCAAAUGGGAGAGACCCUtt-3′ (SEQ ID NO: 3366) 3′-AAAUUUCCGUUUACCCUCUCUGGGAAA-5′ (SEQ ID NO: 3381) AAT-927 Target: 5′-TTTAAAGGCAAATGGGAGAGACCCTTT-3′ (SEQ ID NO: 3396) 5′-AGAAGCUGUCCAGCUGGGUGCUGct-3′ (SEQ ID NO: 3367) 3′-AUUCUUCGACAGGUCGACCCACGACGA-5′ (SEQ ID NO: 3382) AAT-1055 Target: 5′-TAAGAAGCTGTCCAGCTGGGTGCTGCT-3′ (SEQ ID NO: 3397) 5′-AUGCCACCGCCAUCUUCUUCCUGcc-3′ (SEQ ID NO: 3368) 3′-GUUACGGUGGCGGUAGAAGAAGGACGG-5′ (SEQ ID NO: 3383) AAT-1097 Target: 5′-CAATGCCACCGCCATCTTCTTCCTGCC-3′ (SEQ ID NO: 3398) 5′-GCCACCGCCAUCUUCUUCCUGCCtg-3′ (SEQ ID NO: 3369) 3′-UACGGUGGCGGUAGAAGAAGGACGGAC-5′ (SEQ ID NO: 3384) AAT-1099 Target: 5′-ATGCCACCGCCATCTTCTTCCTGCCTG-3′ (SEQ ID NO: 3399) 5′-AGGAGGCACCCCUGAAGCUCUCCaa-3′ (SEQ ID NO: 3370) 3′-UCUCCUCCGUGGGGACUUCGAGAGGUU-5′ (SEQ ID NO: 3385) AAT-1325 Target: 5′-AGAGGAGGCACCCCTGAAGCTCTCCAA-3′ (SEQ ID NO: 3400) 5′-AAGGCCGUGCAUAAGGCUGUGCUga-3′ (SEQ ID NO: 3371) 3′-GGUUCCGGCACGUAUUCCGACACGACU-5′ (SEQ ID NO: 3386) AAT-1348 Target: 5′-CCAAGGCCGTGCATAAGGCTGTGCTGA-3′ (SEQ ID NO: 3401) 5′-CCGUGCAUAAGGCUGUGCUGACCat-3′ (SEQ ID NO: 3372) 3′-CCGGCACGUAUUCCGACACGACUGGUA-5′ (SEQ ID NO: 3387) AAT-1352 Target: 5′-GGCCGTGCATAAGGCTGTGCTGACCAT-3′ (SEQ ID NO: 3402)

TABLE 18 Other Human Anti-a-1 antitrypsin DsiRNAs, Unmodified Duplexes (Asymmetrics) 5′-CCCCAGGGAGAUGCUGCCCAGAAGA-3′ (SEQ ID NO: 3403) 3′-UAGGGGUCCCUCUACGACGGGUCUUCU-5′ (SEQ ID NO: 3373) AAT-364 Target: 5′-ATCCCCAGGGAGATGCTGCCCAGAAGA-3′ (SEQ ID NO: 3388) 5′-GCCUUUGCAAUGCUCUCCCUGGGGA-3′ (SEQ ID NO: 3404) 3′-GUCGGAAACGUUACGAGAGGGACCCCU-5′ (SEQ ID NO: 3374) AAT-535 Target: 5′-CAGCCTTTGCAATGCTCTCCCTGGGGA-3′ (SEQ ID NO: 3389) 5′-GCAAUGCUCUCCCUGGGGACCAAGG-3′ (SEQ ID NO: 3405) 3′-AACGUUACGAGAGGGACCCCUGGUUCC-5′ (SEQ ID NO: 3375) AAT-541 Target: 5′-TTGCAATGCTCTCCCTGGGGACCAAGG-3′ (SEQ ID NO: 3390) 5′-GGGGACCAAGGCUGACACUCACGAU-3′ (SEQ ID NO: 3406) 3′-GACCCCUGGUUCCGACUGUGAGUGCUA-5′ (SEQ ID NO: 3376) AAT-555 Target: 5′-CTGGGGACCAAGGCTGACACTCACGAT-3′ (SEQ ID NO: 3391) 5′-UCCUGGAGGGCCUGAAUUUCAACCU-3′ (SEQ ID NO: 3407) 3′-UUAGGACCUCCCGGACUUAAAGUUGGA-5′ (SEQ ID NO: 3377) AAT-584 Target: 5′-AATCCTGGAGGGCCTGAATTTCAACCT-3′ (SEQ ID NO: 3392) 5′-CAGACAGCCAGCUCCAGCUGACCAC-3′ (SEQ ID NO: 3408) 3′-CGGUCUGUCGGUCGAGGUCGACUGGUG-5′ (SEQ ID NO: 3378) AAT-674 Target: 5′-GCCAGACAGCCAGCTCCAGCTGACCAC-3′ (SEQ ID NO: 3393) 5′-AUCUUCUUUAAAGGCAAAUGGGAGA-3′ (SEQ ID NO: 3409) 3′-UGUAGAAGAAAUUUCCGUUUACCCUCU-5′ (SEQ ID NO: 3379) AAT-919 Target: 5′-ACATCTTCTTTAAAGGCAAATGGGAGA-3′ (SEQ ID NO: 3394) 5′-UUAAAGGCAAAUGGGAGAGACCCUU-3′ (SEQ ID NO: 3410) 3′-GAAAUUUCCGUUUACCCUCUCUGGGAA-5′ (SEQ ID NO: 3380) AAT-926 Target: 5′-CTTTAAAGGCAAATGGGAGAGACCCTT-3′ (SEQ ID NO: 3395) 5′-UAAAGGCAAAUGGGAGAGACCCUUU-3′ (SEQ ID NO: 3411) 3′-AAAUUUCCGUUUACCCUCUCUGGGAAA-5′ (SEQ ID NO: 3381) AAT-927 Target: 5′-TTTAAAGGCAAATGGGAGAGACCCTTT-3′ (SEQ ID NO: 3396) 5′-AGAAGCUGUCCAGCUGGGUGCUGCU-3′ (SEQ ID NO: 3412) 3′-AUUCUUCGACAGGUCGACCCACGACGA-5′ (SEQ ID NO: 3382) AAT-1055 Target: 5′-TAAGAAGCTGTCCAGCTGGGTGCTGCT-3′ (SEQ ID NO: 3397) 5′-AUGCCACCGCCAUCUUCUUCCUGCC-3′ (SEQ ID NO: 3413) 3′-GUUACGGUGGCGGUAGAAGAAGGACGG-5′ (SEQ ID NO: 3383) AAT-1097 Target: 5′-CAATGCCACCGCCATCTTCTTCCTGCC-3′ (SEQ ID NO: 3398) 5′-GCCACCGCCAUCUUCUUCCUGCCUG-3′ (SEQ ID NO: 3414) 3′-UACGGUGGCGGUAGAAGAAGGACGGAC-5′ (SEQ ID NO: 3384) AAT-1099 Target: 5′-ATGCCACCGCCATCTTCTTCCTGCCTG-3′ (SEQ ID NO: 3399) 5′-AGGAGGCACCCCUGAAGCUCUCCAA-3′ (SEQ ID NO: 3415) 3′-UCUCCUCCGUGGGGACUUCGAGAGGUU-5′ (SEQ ID NO: 3385) AAT-1325 Target: 5′-AGAGGAGGCACCCCTGAAGCTCTCCAA-3′ (SEQ ID NO: 3400) 5′-AAGGCCGUGCAUAAGGCUGUGCUGA-3′ (SEQ ID NO: 3416) 3′-GGUUCCGGCACGUAUUCCGACACGACU-5′ (SEQ ID NO: 3386) AAT-1348 Target: 5′-CCAAGGCCGTGCATAAGGCTGTGCTGA-3′ (SEQ ID NO: 3401) 5′-CCGUGCAUAAGGCUGUGCUGACCAU-3′ (SEQ ID NO: 3417) 3′-CCGGCACGUAUUCCGACACGACUGGUA-5′ (SEQ ID NO: 3387) AAT-1352 Target: 5′-GGCCGTGCATAAGGCTGTGCTGACCAT-3′ (SEQ ID NO: 3402)

TABLE 19  Other DsiRNA Target Sequences (21mers) in α-1 antitrypsin mRNA AAT-364 21 nt Target: 5′-AUCCCCAGGGAGAUGCUGCCC-3′(SEQ ID NO: 3418)  AAT-535 21 nt Target: 5′-CAGCCUUUGCAAUGCUCUCCC-3′ (SEQ ID NO: 3419)  AAT-541 21 nt Target: 5′-UUGCAAUGCUCUCCCUGGGGA-3′ (SEQ ID NO: 3420)  AAT-555 21 nt Target: 5′-CUGGGGACCAAGGCUGACACU-3′ (SEQ ID NO: 3421)  AAT-584 21 nt Target: 5′-AAUCCUGGAGGGCCUGAAUUU-3′ (SEQ ID NO: 3422)  AAT-674 21 nt Target: 5′-GCCAGACAGCCAGCUCCAGCU-3′ (SEQ ID NO: 3423)  AAT-919 21 nt Target: 5′-ACAUCUUCUUUAAAGGCAAAU-3′ (SEQ ID NO: 3424)  AAT-926 21 nt Target: 5′-CUUUAAAGGCAAAUGGGAGAG-3′ (SEQ ID NO: 3425)  AAT-927 21 nt Target: 5′-UUUAAAGGCAAAUGGGAGAGA-3′ (SEQ ID NO: 3426)  AAT-1055 21 nt Target: 5′-UAAGAAGCUGUCCAGCUGGGU-3′ (SEQ ID NO: 3427)  AAT-1097 21 nt Target: 5′-CAAUGCCACCGCCAUCUUCUU-3′ (SEQ ID NO: 3428)  AAT-1099 21 nt Target: 5′-AUGCCACCGCCAUCUUCUUCC-3′ (SEQ ID NO: 3429)  AAT-1325 21 nt Target: 5′-AGAGGAGGCACCCCUGAAGCU-3′ (SEQ ID NO: 3430)  AAT-1348 21 nt Target: 5′-CCAAGGCCGUGCAUAAGGCUG-3′ (SEQ ID NO: 3431)  AAT-1352 21 nt Target: 5′-GGCCGUGCAUAAGGCUGUGCU-3′ (SEQ ID NO: 3432)

TABLE 20 Other Human Anti-α-1 antitrypsin “Blunt/Blunt” DsiRNAs 5′-AUCCCCAGGGAGAUGCUGCCCAGAAGA-3′ (SEQ ID NO: 3433) 3′-UAGGGGUCCCUCUACGACGGGUCUUCU-5′ (SEQ ID NO: 3373) AAT-364 Target: 5′-ATCCCCAGGGAGATGCTGCCCAGAAGA-3′ (SEQ ID NO: 3388) 5′-CAGCCUUUGCAAUGCUCUCCCUGGGGA-3′ (SEQ ID NO: 3434) 3′-GUCGGAAACGUUACGAGAGGGACCCCU-5′ (SEQ ID NO: 3374) AAT-535 Target: 5′-CAGCCTTTGCAATGCTCTCCCTGGGGA-3′ (SEQ ID NO: 3389) 5′-UUGCAAUGCUCUCCCUGGGGACCAAGG-3′ (SEQ ID NO: 3435) 3′-AACGUUACGAGAGGGACCCCUGGUUCC-5′ (SEQ ID NO: 3375) AAT-541 Target: 5′-TTGCAATGCTCTCCCTGGGGACCAAGG-3′ (SEQ ID NO: 3390) 5′-CUGGGGACCAAGGCUGACACUCACGAU-3′ (SEQ ID NO: 3436) 3′-GACCCCUGGUUCCGACUGUGAGUGCUA-5′ (SEQ ID NO: 3376) AAT-555 Target: 5′-CTGGGGACCAAGGCTGACACTCACGAT-3′ (SEQ ID NO: 3391) 5′-AAUCCUGGAGGGCCUGAAUUUCAACCU-3′ (SEQ ID NO: 3437) 3′-UUAGGACCUCCCGGACUUAAAGUUGGA-5′ (SEQ ID NO: 3377) AAT-584 Target: 5′-AATCCTGGAGGGCCTGAATTTCAACCT-3′ (SEQ ID NO: 3392) 5′-GCCAGACAGCCAGCUCCAGCUGACCAC-3′ (SEQ ID NO: 3438) 3′-CGGUCUGUCGGUCGAGGUCGACUGGUG-5′ (SEQ ID NO: 3378) AAT-674 Target: 5′-GCCAGACAGCCAGCTCCAGCTGACCAC-3′ (SEQ ID NO: 3393) 5′-ACAUCUUCUUUAAAGGCAAAUGGGAGA-3′ (SEQ ID NO: 3439) 3′-UGUAGAAGAAAUUUCCGUUUACCCUCU-5′ (SEQ ID NO: 3379) AAT-919 Target: 5′-ACATCTTCTTTAAAGGCAAATGGGAGA-3′ (SEQ ID NO: 3394) 5′-CUUUAAAGGCAAAUGGGAGAGACCCUU-3′ (SEQ ID NO: 3440) 3′-GAAAUUUCCGUUUACCCUCUCUGGGAA-5′ (SEQ ID NO: 3380) AAT-926 Target: 5′-CTTTAAAGGCAAATGGGAGAGACCCTT-3′ (SEQ ID NO: 3395) 5′-UUUAAAGGCAAAUGGGAGAGACCCUUU-3′ (SEQ ID NO: 3441) 3′-AAAUUUCCGUUUACCCUCUCUGGGAAA-5′ (SEQ ID NO: 3381) AAT-927 Target: 5′-TTTAAAGGCAAATGGGAGAGACCCTTT-3′ (SEQ ID NO: 3396) 5′-UAAGAAGCUGUCCAGCUGGGUGCUGCU-3′ (SEQ ID NO: 3442) 3′-AUUCUUCGACAGGUCGACCCACGACGA-5′ (SEQ ID NO: 3382) AAT-1055 Target: 5′-TAAGAAGCTGTCCAGCTGGGTGCTGCT-3′ (SEQ ID NO: 3397) 5′-CAAUGCCACCGCCAUCUUCUUCCUGCC-3′ (SEQ ID NO: 3443) 3′-GUUACGGUGGCGGUAGAAGAAGGACGG-5′ (SEQ ID NO: 3383) AAT-1097 Target: 5′-CAATGCCACCGCCATCTTCTTCCTGCC-3′ (SEQ ID NO: 3398) 5′-AUGCCACCGCCAUCUUCUUCCUGCCUG-3′ (SEQ ID NO: 3444) 3′-UACGGUGGCGGUAGAAGAAGGACGGAC-5′ (SEQ ID NO: 3384) AAT-1099 Target: 5′-ATGCCACCGCCATCTTCTTCCTGCCTG-3′ (SEQ ID NO: 3399) 5′-AGAGGAGGCACCCCUGAAGCUCUCCAA-3′ (SEQ ID NO: 3445) 3′-UCUCCUCCGUGGGGACUUCGAGAGGUU-5′ (SEQ ID NO: 3385) AAT-1325 Target: 5′-AGAGGAGGCACCCCTGAAGCTCTCCAA-3′ (SEQ ID NO: 3400) 5′-CCAAGGCCGUGCAUAAGGCUGUGCUGA-3′ (SEQ ID NO: 3446) 3′-GGUUCCGGCACGUAUUCCGACACGACU-5′ (SEQ ID NO: 3386) AAT-1348 Target: 5′-CCAAGGCCGTGCATAAGGCTGTGCTGA-3′ (SEQ ID NO: 3401) 5′-GGCCGUGCAUAAGGCUGUGCUGACCAU-3′ (SEQ ID NO: 3447) 3′-CCGGCACGUAUUCCGACACGACUGGUA-5′ (SEQ ID NO: 3387) AAT-1352 Target: 5′-GGCCGTGCATAAGGCTGTGCTGACCAT-3′ (SEQ ID NO: 3402)

TABLE 21 Other DsiRNA Component 19 Nucleotide Target Sequences in α-1 antitrypsin mRNA AAT-364 19 nt Target #1: 5′-CCCCAGGGAGAUGCUGCCC-3′ (SEQ ID NO: 3448) AAT-364 19 nt Target #2: 5′-UCCCCAGGGAGAUGCUGCC-3′ (SEQ ID NO: 3463) AAT-364 19 nt Target #3: 5′-AUCCCCAGGGAGAUGCUGC-3′ (SEQ ID NO: 3478) AAT-535 19 nt Target #1: 5′-GCCUUUGCAAUGCUCUCCC-3′ (SEQ ID NO: 3449) AAT-535 19 nt Target #2: 5′-AGCCUUUGCAAUGCUCUCC-3′ (SEQ ID NO: 3464) AAT-535 19 nt Target #3: 5′-CAGCCUUUGCAAUGCUCUC-3′ (SEQ ID NO: 3479) AAT-541 19 nt Target #1: 5′-GCAAUGCUCUCCCUGGGGA-3′ (SEQ ID NO: 3450) AAT-541 19 nt Target #2: 5′-UGCAAUGCUCUCCCUGGGG-3′ (SEQ ID NO: 3465) AAT-541 19 nt Target #3: 5′-UUGCAAUGCUCUCCCUGGG-3′ (SEQ ID NO: 3480) AAT-555 19 nt Target #1: 5′-GGGGACCAAGGCUGACACU-3′ (SEQ ID NO: 3451) AAT-555 19 nt Target #2: 5′-UGGGGACCAAGGCUGACAC-3′ (SEQ ID NO: 3466) AAT-555 19 nt Target #3: 5′-CUGGGGACCAAGGCUGACA-3′ (SEQ ID NO: 3481) AAT-584 19 nt Target #1: 5′-UCCUGGAGGGCCUGAAUUU-3′ (SEQ ID NO: 3452) AAT-584 19 nt Target #2: 5′-AUCCUGGAGGGCCUGAAUU-3′ (SEQ ID NO: 3467) AAT-584 19 nt Target #3: 5′-AAUCCUGGAGGGCCUGAAU-3′ (SEQ ID NO: 3482) AAT-674 19 nt Target #1: 5′-CAGACAGCCAGCUCCAGCU-3′ (SEQ ID NO: 3453) AAT-674 19 nt Target #2: 5′-CCAGACAGCCAGCUCCAGC-3′ (SEQ ID NO: 3468) AAT-674 19 nt Target #3: 5′-GCCAGACAGCCAGCUCCAG-3′ (SEQ ID NO: 3483) AAT-919 19 nt Target #1: 5′-AUCUUCUUUAAAGGCAAAU-3′ (SEQ ID NO: 3454) AAT-919 19 nt Target #2: 5′-CAUCUUCUUUAAAGGCAAA-3′ (SEQ ID NO: 3469) AAT-919 19 nt Target #3: 5′-ACAUCUUCUUUAAAGGCAA-3′ (SEQ ID NO: 3484) AAT-926 19 nt Target #1: 5′-UUAAAGGCAAAUGGGAGAG-3′ (SEQ ID NO: 3455) AAT-926 19 nt Target #2: 5′-UUUAAAGGCAAAUGGGAGA-3′ (SEQ ID NO: 3470) AAT-926 19 nt Target #3: 5′-CUUUAAAGGCAAAUGGGAG-3′ (SEQ ID NO: 3485) AAT-927 19 nt Target #1: 5′-UAAAGGCAAAUGGGAGAGA-3′ (SEQ ID NO: 3456) AAT-927 19 nt Target #2: 5′-UUAAAGGCAAAUGGGAGAG-3′ (SEQ ID NO: 3471) AAT-927 19 nt Target #3: 5′-UUUAAAGGCAAAUGGGAGA-3′ (SEQ ID NO: 3486) AAT-1055 19 nt Target #1: 5′-AGAAGCUGUCCAGCUGGGU-3′ (SEQ ID NO: 3457) AAT-1055 19 nt Target #2: 5′-AAGAAGCUGUCCAGCUGGG-3′ (SEQ ID NO: 3472) AAT-1055 19 nt Target #3: 5′-UAAGAAGCUGUCCAGCUGG-3′ (SEQ ID NO: 3487) AAT-1097 19 nt Target #1: 5′-AUGCCACCGCCAUCUUCUU-3′ (SEQ ID NO: 3458) AAT-1097 19 nt Target #2: 5′-AAUGCCACCGCCAUCUUCU-3′ (SEQ ID NO: 3473) AAT-1097 19 nt Target #3: 5′-CAAUGCCACCGCCAUCUUC-3′ (SEQ ID NO: 3488) AAT-1099 19 nt Target #1: 5′-GCCACCGCCAUCUUCUUCC-3′ (SEQ ID NO: 3459) AAT-1099 19 nt Target #2: 5′-UGCCACCGCCAUCUUCUUC-3′ (SEQ ID NO: 3474) AAT-1099 19 nt Target #3: 5′-AUGCCACCGCCAUCUUCUU-3′ (SEQ ID NO: 3489) AAT-1325 19 nt Target #1: 5′-AGGAGGCACCCCUGAAGCU-3′ (SEQ ID NO: 3460) AAT-1325 19 nt Target #2: 5′-GAGGAGGCACCCCUGAAGC-3′ (SEQ ID NO: 3475) AAT-1325 19 nt Target #3: 5′-AGAGGAGGCACCCCUGAAG-3′ (SEQ ID NO: 3490) AAT-1348 19 nt Target #1: 5′-AAGGCCGUGCAUAAGGCUG-3′ (SEQ ID NO: 3461) AAT-1348 19 nt Target #2: 5′-CAAGGCCGUGCAUAAGGCU-3′ (SEQ ID NO: 3476) AAT-1348 19 nt Target #3: 5′-CCAAGGCCGUGCAUAAGGC-3′ (SEQ ID NO: 3491) AAT-1352 19 nt Target #1: 5′-CCGUGCAUAAGGCUGUGCU-3′ (SEQ ID NO: 3462) AAT-1352 19 nt Target #2: 5′-GCCGUGCAUAAGGCUGUGC-3′ (SEQ ID NO: 3477) AAT-1352 19 nt Target #3: 5′-GGCCGUGCAUAAGGCUGUG-3′ (SEQ ID NO: 3492)

Within Tables 2, 3, 5, 7, 8, 10, 12, 13, 15, 17, 18 and 20 above, underlined residues indicate 2′-O-methyl residues, UPPER CASE indicates ribonucleotides, and lower case denotes deoxyribonucleotides. The DsiRNA agents of Tables 2-3, 7-8, 12-13 and 17-18 above are 25/27mer agents possessing a blunt end. The structures and/or modification patterning of the agents of Tables 2-3, 7-8, 12-13 and 17-18 above can be readily adapted to the above generic sequence structures, e.g., the 3′ overhang of the second strand can be extended or contracted, 2′-O-methylation of the second strand can be expanded towards the 5′ end of the second strand, optionally at alternating sites, etc. Such further modifications are optional, as 25/27mer DsiRNAs with such modifications can also be readily designed from the above DsiRNA agents and are also expected to be functional inhibitors of α-1 antitrypsin expression. Similarly, the 27mer “blunt/blunt” DsiRNA structures and/or modification patterns of the agents of Tables 5, 10, 15 and 20 above can also be readily adapted to the above generic sequence structures, e.g., for application of modification patterning of the antisense strand to such structures and/or adaptation of such sequences to the above generic structures.

In certain embodiments, 27mer DsiRNAs possessing independent strand lengths each of 27 nucleotides are designed and synthesized for targeting of the same sites within the α-1 antitrypsin transcript as the asymmetric “25/27” structures shown in Tables 2-3, 7-8, 12-13 and 17-18 herein. Exemplary “27/27” DsiRNAs are optionally designed with a “blunt/blunt” structure as shown for the DsiRNAs of Tables 5, 10, 15 and 20 above.

In certain embodiments, the dsRNA agents of the invention require, e.g., at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25 or at least 26 residues of the first strand to be complementary to corresponding residues of the second strand. In certain related embodiments, these first strand residues complementary to corresponding residues of the second strand are optionally consecutive residues.

In certain DsiRNAmm (“DsiRNA mismatch”) embodiments of the instant invention, mismatched base pairs are located within a “mismatch-tolerant region” which is defined herein with respect to the location of the projected Ago2 cut site of the corresponding target nucleic acid. The mismatch tolerant region is located “upstream of” the projected Ago2 cut site of the target strand. “Upstream” in this context will be understood as the 5′-most portion of the DsiRNAmm duplex, where 5′ refers to the orientation of the sense strand of the DsiRNA duplex. Therefore, the mismatch tolerant region is upstream of the base on the sense (passenger) strand that corresponds to the projected Ago2 cut site of the target nucleic acid (see FIG. 1); alternatively, when referring to the antisense (guide) strand of the DsiRNAmm, the mismatch tolerant region can also be described as positioned downstream of the base that is complementary to the projected Ago2 cut site of the target nucleic acid, that is, the 3′-most portion of the antisense strand of the DsiRNAmm (where position 1 of the antisense strand is the 5′ terminal nucleotide of the antisense strand, see FIG. 1).

In one embodiment, for example with numbering as depicted in FIG. 1, the mismatch tolerant region is positioned between and including base pairs 3-9 when numbered from the nucleotide starting at the 5′ end of the sense strand of the duplex. Therefore, a DsiRNAmm of the invention possesses a single mismatched base pair at any one of positions 3, 4, 5, 6, 7, 8 or 9 of the sense strand of a right-hand extended DsiRNA (where position 1 is the 5′ terminal nucleotide of the sense strand and position 9 is the nucleotide residue of the sense strand that is immediately 5′ of the projected Ago2 cut site of the target α-1 antitrypsin RNA sequence corresponding to the sense strand sequence). In certain embodiments, for a DsiRNAmm that possesses a mismatched base pair nucleotide at any of positions 3, 4, 5, 6, 7, 8 or 9 of the sense strand, the corresponding mismatched base pair nucleotide of the antisense strand not only forms a mismatched base pair with the DsiRNAmm sense strand sequence, but also forms a mismatched base pair with a DsiRNAmm target α-1 antitrypsin RNA sequence (thus, complementarity between the antisense strand sequence and the sense strand sequence is disrupted at the mismatched base pair within the DsiRNAmm, and complementarity is similarly disrupted between the antisense strand sequence of the DsiRNAmm and the target α-1 antitrypsin RNA sequence). In alternative embodiments, the mismatch base pair nucleotide of the antisense strand of a DsiRNAmm only form a mismatched base pair with a corresponding nucleotide of the sense strand sequence of the DsiRNAmm, yet base pairs with its corresponding target α-1 antitrypsin RNA sequence nucleotide (thus, complementarity between the antisense strand sequence and the sense strand sequence is disrupted at the mismatched base pair within the DsiRNAmm, yet complementarity is maintained between the antisense strand sequence of the DsiRNAmm and the target α-1 antitrypsin RNA sequence).

A DsiRNAmm of the invention that possesses a single mismatched base pair within the mismatch-tolerant region (mismatch region) as described above (e.g., a DsiRNAmm harboring a mismatched nucleotide residue at any one of positions 3, 4, 5, 6, 7, 8 or 9 of the sense strand) can further include one, two or even three additional mismatched base pairs. In preferred embodiments, these one, two or three additional mismatched base pairs of the DsiRNAmm occur at position(s) 3, 4, 5, 6, 7, 8 and/or 9 of the sense strand (and at corresponding residues of the antisense strand). In one embodiment where one additional mismatched base pair is present within a DsiRNAmm, the two mismatched base pairs of the sense strand can occur, e.g., at nucleotides of both position 4 and position 6 of the sense strand (with mismatch also occurring at corresponding nucleotide residues of the antisense strand).

In DsiRNAmm agents possessing two mismatched base pairs, mismatches can occur consecutively (e.g., at consecutive positions along the sense strand nucleotide sequence). Alternatively, nucleotides of the sense strand that form mismatched base pairs with the antisense strand sequence can be interspersed by nucleotides that base pair with the antisense strand sequence (e.g., for a DsiRNAmm possessing mismatched nucleotides at positions 3 and 6, but not at positions 4 and 5, the mismatched residues of sense strand positions 3 and 6 are interspersed by two nucleotides that form matched base pairs with corresponding residues of the antisense strand). For example, two residues of the sense strand (located within the mismatch-tolerant region of the sense strand) that form mismatched base pairs with the corresponding antisense strand sequence can occur with zero, one, two, three, four or five matched base pairs located between these mismatched base pairs.

For certain DsiRNAmm agents possessing three mismatched base pairs, mismatches can occur consecutively (e.g., in a triplet along the sense strand nucleotide sequence). Alternatively, nucleotides of the sense strand that form mismatched base pairs with the antisense strand sequence can be interspersed by nucleotides that form matched base pairs with the antisense strand sequence (e.g., for a DsiRNAmm possessing mismatched nucleotides at positions 3, 4 and 8, but not at positions 5, 6 and 7, the mismatched residues of sense strand positions 3 and 4 are adjacent to one another, while the mismatched residues of sense strand positions 4 and 8 are interspersed by three nucleotides that form matched base pairs with corresponding residues of the antisense strand). For example, three residues of the sense strand (located within the mismatch-tolerant region of the sense strand) that form mismatched base pairs with the corresponding antisense strand sequence can occur with zero, one, two, three or four matched base pairs located between any two of these mismatched base pairs.

For certain DsiRNAmm agents possessing four mismatched base pairs, mismatches can occur consecutively (e.g., in a quadruplet along the sense strand nucleotide sequence). Alternatively, nucleotides of the sense strand that form mismatched base pairs with the antisense strand sequence can be interspersed by nucleotides that form matched base pairs with the antisense strand sequence (e.g., for a DsiRNAmm possessing mismatched nucleotides at positions 3, 5, 7 and 8, but not at positions 4 and 6, the mismatched residues of sense strand positions 7 and 8 are adjacent to one another, while the mismatched residues of sense strand positions 3 and 5 are interspersed by one nucleotide that forms a matched base pair with the corresponding residue of the antisense strand—similarly, the mismatched residues of sense strand positions 5 and 7 are also interspersed by one nucleotide that forms a matched base pair with the corresponding residue of the antisense strand). For example, four residues of the sense strand (located within the mismatch-tolerant region of the sense strand) that form mismatched base pairs with the corresponding antisense strand sequence can occur with zero, one, two or three matched base pairs located between any two of these mismatched base pairs.

In another embodiment, for example with numbering also as depicted in FIG. 1, a DsiRNAmm of the invention comprises a mismatch tolerant region which possesses a single mismatched base pair nucleotide at any one of positions 17, 18, 19, 20, 21, 22 or 23 of the antisense strand of the DsiRNA (where position 1 is the 5′ terminal nucleotide of the antisense strand and position 17 is the nucleotide residue of the antisense strand that is immediately 3′ (downstream) in the antisense strand of the projected Ago2 cut site of the target α-1 antitrypsin RNA sequence sufficiently complementary to the antisense strand sequence). In certain embodiments, for a DsiRNAmm that possesses a mismatched base pair nucleotide at any of positions 17, 18, 19, 20, 21, 22 or 23 of the antisense strand with respect to the sense strand of the DsiRNAmm, the mismatched base pair nucleotide of the antisense strand not only forms a mismatched base pair with the DsiRNAmm sense strand sequence, but also forms a mismatched base pair with a DsiRNAmm target α-1 antitrypsin RNA sequence (thus, complementarity between the antisense strand sequence and the sense strand sequence is disrupted at the mismatched base pair within the DsiRNAmm, and complementarity is similarly disrupted between the antisense strand sequence of the DsiRNAmm and the target α-1 antitrypsin RNA sequence). In alternative embodiments, the mismatch base pair nucleotide of the antisense strand of a DsiRNAmm only forms a mismatched base pair with a corresponding nucleotide of the sense strand sequence of the DsiRNAmm, yet base pairs with its corresponding target α-1 antitrypsin RNA sequence nucleotide (thus, complementarity between the antisense strand sequence and the sense strand sequence is disrupted at the mismatched base pair within the DsiRNAmm, yet complementarity is maintained between the antisense strand sequence of the DsiRNAmm and the target α-1 antitrypsin RNA sequence).

A DsiRNAmm of the invention that possesses a single mismatched base pair within the mismatch-tolerant region as described above (e.g., a DsiRNAmm harboring a mismatched nucleotide residue at positions 17, 18, 19, 20, 21, 22 or 23 of the antisense strand) can further include one, two or even three additional mismatched base pairs. In preferred embodiments, these one, two or three additional mismatched base pairs of the DsiRNAmm occur at position(s) 17, 18, 19, 20, 21, 22 and/or 23 of the antisense strand (and at corresponding residues of the sense strand). In one embodiment where one additional mismatched base pair is present within a DsiRNAmm, the two mismatched base pairs of the antisense strand can occur, e.g., at nucleotides of both position 18 and position 20 of the antisense strand (with mismatch also occurring at corresponding nucleotide residues of the sense strand).

In DsiRNAmm agents possessing two mismatched base pairs, mismatches can occur consecutively (e.g., at consecutive positions along the antisense strand nucleotide sequence). Alternatively, nucleotides of the antisense strand that form mismatched base pairs with the sense strand sequence can be interspersed by nucleotides that base pair with the sense strand sequence (e.g., for a DsiRNAmm possessing mismatched nucleotides at positions 17 and 20, but not at positions 18 and 19, the mismatched residues of antisense strand positions 17 and 20 are interspersed by two nucleotides that form matched base pairs with corresponding residues of the sense strand). For example, two residues of the antisense strand (located within the mismatch-tolerant region of the sense strand) that form mismatched base pairs with the corresponding sense strand sequence can occur with zero, one, two, three, four, five, six or seven matched base pairs located between these mismatched base pairs.

For certain DsiRNAmm agents possessing three mismatched base pairs, mismatches can occur consecutively (e.g., in a triplet along the antisense strand nucleotide sequence). Alternatively, nucleotides of the antisense strand that form mismatched base pairs with the sense strand sequence can be interspersed by nucleotides that form matched base pairs with the sense strand sequence (e.g., for a DsiRNAmm possessing mismatched nucleotides at positions 17, 18 and 22, but not at positions 19, 20 and 21, the mismatched residues of antisense strand positions 17 and 18 are adjacent to one another, while the mismatched residues of antisense strand positions 18 and 122 are interspersed by three nucleotides that form matched base pairs with corresponding residues of the sense strand). For example, three residues of the antisense strand (located within the mismatch-tolerant region of the antisense strand) that form mismatched base pairs with the corresponding sense strand sequence can occur with zero, one, two, three, four, five or six matched base pairs located between any two of these mismatched base pairs.

For certain DsiRNAmm agents possessing four mismatched base pairs, mismatches can occur consecutively (e.g., in a quadruplet along the antisense strand nucleotide sequence). Alternatively, nucleotides of the antisense strand that form mismatched base pairs with the sense strand sequence can be interspersed by nucleotides that form matched base pairs with the sense strand sequence (e.g., for a DsiRNAmm possessing mismatched nucleotides at positions 18, 20, 22 and 23, but not at positions 19 and 21, the mismatched residues of antisense strand positions 22 and 23 are adjacent to one another, while the mismatched residues of antisense strand positions 18 and 20 are interspersed by one nucleotide that forms a matched base pair with the corresponding residue of the sense strand—similarly, the mismatched residues of antisense strand positions 20 and 22 are also interspersed by one nucleotide that forms a matched base pair with the corresponding residue of the sense strand). For example, four residues of the antisense strand (located within the mismatch-tolerant region of the antisense strand) that form mismatched base pairs with the corresponding sense strand sequence can occur with zero, one, two, three, four or five matched base pairs located between any two of these mismatched base pairs.

For reasons of clarity, the location(s) of mismatched nucleotide residues within the above DsiRNAmm agents are numbered in reference to the 5′ terminal residue of either sense or antisense strands of the DsiRNAmm. The numbering of positions located within the mismatch-tolerant region (mismatch region) of the antisense strand can shift with variations in the proximity of the 5′ terminus of the sense or antisense strand to the projected Ago2 cleavage site. Thus, the location(s) of preferred mismatch sites within either antisense strand or sense strand can also be identified as the permissible proximity of such mismatches to the projected Ago2 cut site. Accordingly, in one preferred embodiment, the position of a mismatch nucleotide of the sense strand of a DsiRNAmm is the nucleotide residue of the sense strand that is located immediately 5′ (upstream) of the projected Ago2 cleavage site of the corresponding target α-1 antitrypsin RNA sequence. In other preferred embodiments, a mismatch nucleotide of the sense strand of a DsiRNAmm is positioned at the nucleotide residue of the sense strand that is located two nucleotides 5′ (upstream) of the projected Ago2 cleavage site, three nucleotides 5′ (upstream) of the projected Ago2 cleavage site, four nucleotides 5′ (upstream) of the projected Ago2 cleavage site, five nucleotides 5′ (upstream) of the projected Ago2 cleavage site, six nucleotides 5′ (upstream) of the projected Ago2 cleavage site, seven nucleotides 5′ (upstream) of the projected Ago2 cleavage site, eight nucleotides 5′ (upstream) of the projected Ago2 cleavage site, or nine nucleotides 5′ (upstream) of the projected Ago2 cleavage site.

Exemplary single mismatch-containing 25/27mer DsiRNAs (DsiRNAmm) include the following structures (such mismatch-containing structures may also be incorporated into other exemplary DsiRNA structures shown herein).

5′-XX^(M)XXXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXX_(M)XXXXXXXXXXXXXXXXXXXXXX-5′ 5′-XXX^(M)XXXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXX_(M)XXXXXXXXXXXXXXXXXXXXX-5′ 5′-XXXX^(M)XXXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXX_(M)XXXXXXXXXXXXXXXXXXXX-5′ 5′-XXXXX^(M)XXXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXX_(M)XXXXXXXXXXXXXXXXXXX-5′ 5′-XXXXXX^(M)XXXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXX_(M)XXXXXXXXXXXXXXXXXX-5′ 5′-XXXXXXX^(M)XXXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXX_(M)XXXXXXXXXXXXXXXXX-5′ 5′-XXXXXXXX^(M)XXXXXXXXXXXXXXDD-3′ 3′-XXXXXXXXXX_(M)XXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “D”=DNA and “M”=Nucleic acid residues (RNA, DNA or non-natural or modified nucleic acids) that do not base pair (hydrogen bond) with corresponding “M” residues of otherwise complementary strand when strands are annealed. Any of the residues of such agents can optionally be 2′-O-methyl RNA monomers—alternating positioning of 2′-O-methyl RNA monomers that commences from the 3′-terminal residue of the bottom (second) strand, as shown above, can also be used in the above DsiRNAmm agents. For the above mismatch structures, the top strand is the sense strand, and the bottom strand is the antisense strand.

In certain embodiments, a DsiRNA of the invention can contain mismatches that exist in reference to the target α-1 antitrypsin RNA sequence yet do not necessarily exist as mismatched base pairs within the two strands of the DsiRNA—thus, a DsiRNA can possess perfect complementarity between first and second strands of a DsiRNA, yet still possess mismatched residues in reference to a target α-1 antitrypsin RNA (which, in certain embodiments, may be advantageous in promoting efficacy and/or potency and/or duration of effect). In certain embodiments, where mismatches occur between antisense strand and target α-1 antitrypsin RNA sequence, the position of a mismatch is located within the antisense strand at a position(s) that corresponds to a sequence of the sense strand located 5′ of the projected Ago2 cut site of the target region—e.g., antisense strand residue(s) positioned within the antisense strand to the 3′ of the antisense residue which is complementary to the projected Ago2 cut site of the target sequence.

Exemplary 25/27mer DsiRNAs that harbor a single mismatched residue in reference to target sequences include the following structures.

Target RNA Sequence: 5′-. . . AXXXXXXXXXXXXXXXXXXXX . . .-3′ DsiRNAmm Sense Strand: 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ DsiRNAmm Antisense Strand: 3′-EXXXXXXXXXXXXXXXXXXXXXXXXXX-5′ Target RNA Sequence: 5′-. . . XAXXXXXXXXXXXXXXXXXXX . . .-3′ DsiRNAmm Sense Strand: 5′-XXXXXXXXXXXXXXXXXXXXXXXDD-3′ DsiRNAmm Antisense Strand: 3′-XEXXXXXXXXXXXXXXXXXXXXXXXXX-5′ Target RNA Sequence: 5′-. . . AXXXXXXXXXXXXXXXXXX . . .-3′ DsiRNAmm Sense Strand: 5′-BXXXXXXXXXXXXXXXXXXXXXXDD-3′ DsiRNAmm Antisense Strand: 3′-XXEXXXXXXXXXXXXXXXXXXXXXXXX-5′ Target RNA Sequence: 5′-. . . XAXXXXXXXXXXXXXXXXX . . .-3′ DsiRNAmm Sense Strand: 5′-XBXXXXXXXXXXXXXXXXXXXXXDD-3′ DsiRNAmm Antisense Strand: 3′-XXXEXXXXXXXXXXXXXXXXXXXXXXX-5′ Target RNA Sequence: 5′-. . . XXAXXXXXXXXXXXXXXXX . . .-3′ DsiRNAmm Sense Strand: 5′-XXBXXXXXXXXXXXXXXXXXXXXDD-3′ DsiRNAmm Antisense Strand: 3′-XXXXEXXXXXXXXXXXXXXXXXXXXXX-5′ Target RNA Sequence: 5′-. . . XXXAXXXXXXXXXXXXXXX . . .-3′ DsiRNAmm Sense Strand: 5′-XXXBXXXXXXXXXXXXXXXXXXXDD-3′ DsiRNAmm Antisense Strand: 3′-XXXXXEXXXXXXXXXXXXXXXXXXXXX-5′ Target RNA Sequence: 5′-. . . XXXXAXXXXXXXXXXXXXX . .  DsiRNAmm Sense Strand: 5′-XXXXBXXXXXXXXXXXXXXXXXXDD-3′ DsiRNAmm Antisense Strand: 3′-XXXXXXEXXXXXXXXXXXXXXXXXXXX-5′ Target RNA Sequence: 5′-. . . XXXXXAXXXXXXXXXXXXX . .  DsiRNAmm Sense Strand: 5′-XXXXXBXXXXXXXXXXXXXXXXXDD-3′ DsiRNAmm Antisense Strand: 3′-XXXXXXXEXXXXXXXXXXXXXXXXXXX-5′ Target RNA Sequence: 5′-. . . XXXXXXAXXXXXXXXXXXX . .  DsiRNAmm Sense Strand: 5′-XXXXXXBXXXXXXXXXXXXXXXXDD-3′ DsiRNAmm Antisense Strand: 3′-XXXXXXXXEXXXXXXXXXXXXXXXXXX-5′ Target RNA Sequence: 5′-. . . XXXXXXXAXXXXXXXXXXX . . .-3′ DsiRNAmm Sense Strand: 5′-XXXXXXXBXXXXXXXXXXXXXXXDD-3′ DsiRNAmm Antisense Strand: 3′-XXXXXXXXXEXXXXXXXXXXXXXXXXX-5′ Target RNA Sequence: 5′-. . . XXXXXXXXAXXXXXXXXXX . . .-3′ DsiRNAmm Sense Strand: 5′-XXXXXXXXBXXXXXXXXXXXXXXDD-3′ DsiRNAmm Antisense Strand: 3′-XXXXXXXXXXEXXXXXXXXXXXXXXXX-5′ wherein “X”=RNA, “D”=DNA and “E”=Nucleic acid residues (RNA, DNA or non-natural or modified nucleic acids) that do not base pair (hydrogen bond) with corresponding “A” RNA residues of otherwise complementary (target) strand when strands are annealed, yet optionally do base pair with corresponding “B” residues (“B” residues are also RNA, DNA or non-natural or modified nucleic acids). Any of the residues of such agents can optionally be 2′-O-methyl RNA monomers—alternating positioning of 2′-O-methyl RNA monomers that commences from the 3′-terminal residue of the bottom (second) strand, as shown above, can also be used in the above DsiRNA agents.

In certain embodiments, the guide strand of a dsRNA of the invention that is sufficiently complementary to a target RNA (e.g., mRNA) along at least 19 nucleotides of the target gene sequence to reduce target gene expression is not perfectly complementary to the at least 19 nucleotide long target gene sequence. Rather, it is appreciated that the guide strand of a dsRNA of the invention that is sufficiently complementary to a target mRNA along at least 19 nucleotides of a target RNA sequence to reduce target gene expression can have one, two, three, or even four or more nucleotides that are mismatched with the 19 nucleotide or longer target strand sequence. Thus, for a 19 nucleotide target RNA sequence, the guide strand of a dsRNA of the invention can be sufficiently complementary to the target RNA sequence to reduce target gene levels while possessing, e.g., only 15/19, 16/19, 17/19 or 18/19 matched nucleotide residues between guide strand and target RNA sequence.

In addition to the above-exemplified structures, dsRNAs of the invention can also possess one, two or three additional residues that form further mismatches with the target α-1 antitrypsin RNA sequence. Such mismatches can be consecutive, or can be interspersed by nucleotides that form matched base pairs with the target α-1 antitryp sin RNA sequence. Where interspersed by nucleotides that form matched base pairs, mismatched residues can be spaced apart from each other within a single strand at an interval of one, two, three, four, five, six, seven or even eight base paired nucleotides between such mismatch-forming residues.

As for the above-described DsiRNAmm agents, a preferred location within dsRNAs (e.g., DsiRNAs) for antisense strand nucleotides that form mismatched base pairs with target α-1 antitrypsin RNA sequence (yet may or may not form mismatches with corresponding sense strand nucleotides) is within the antisense strand region that is located 3′ (downstream) of the antisense strand sequence which is complementary to the projected Ago2 cut site of the DsiRNA (e.g., in FIG. 1, the region of the antisense strand which is 3′ of the projected Ago2 cut site is preferred for mismatch-forming residues and happens to be located at positions 17-23 of the antisense strand for the 25/27mer agent shown in FIG. 1). Thus, in one embodiment, the position of a mismatch nucleotide (in relation to the target α-1 antitrypsin RNA sequence) of the antisense strand of a DsiRNAmm is the nucleotide residue of the antisense strand that is located immediately 3′ (downstream) within the antisense strand sequence of the projected Ago2 cleavage site of the corresponding target α-1 antitrypsin RNA sequence. In other preferred embodiments, a mismatch nucleotide of the antisense strand of a DsiRNAmm (in relation to the target α-1 antitrypsin RNA sequence) is positioned at the nucleotide residue of the antisense strand that is located two nucleotides 3′ (downstream) of the corresponding projected Ago2 cleavage site, three nucleotides 3′ (downstream) of the corresponding projected Ago2 cleavage site, four nucleotides 3′ (downstream) of the corresponding projected Ago2 cleavage site, five nucleotides 3′ (downstream) of the corresponding projected Ago2 cleavage site, six nucleotides 3′ (downstream) of the projected Ago2 cleavage site, seven nucleotides 3′ (downstream) of the projected Ago2 cleavage site, eight nucleotides 3′ (downstream) of the projected Ago2 cleavage site, or nine nucleotides 3′ (downstream) of the projected Ago2 cleavage site.

In dsRNA agents possessing two mismatch-forming nucleotides of the antisense strand (where mismatch-forming nucleotides are mismatch forming in relation to target α-1 antitrypsin RNA sequence), mismatches can occur consecutively (e.g., at consecutive positions along the antisense strand nucleotide sequence). Alternatively, nucleotides of the antisense strand that form mismatched base pairs with the target α-1 antitrypsin RNA sequence can be interspersed by nucleotides that base pair with the target α-1 antitrypsin RNA sequence (e.g., for a DsiRNA possessing mismatch-forming nucleotides at positions 17 and 20 (starting from the 5′ terminus (position 1) of the antisense strand of the 25/27mer agent shown in FIG. 1), but not at positions 18 and 19, the mismatched residues of sense strand positions 17 and 20 are interspersed by two nucleotides that form matched base pairs with corresponding residues of the target α-1 antitrypsin RNA sequence). For example, two residues of the antisense strand (located within the mismatch-tolerant region of the antisense strand) that form mismatched base pairs with the corresponding target α-1 antitrypsin RNA sequence can occur with zero, one, two, three, four or five matched base pairs (with respect to target α-1 antitrypsin RNA sequence) located between these mismatch-forming base pairs.

For certain dsRNAs possessing three mismatch-forming base pairs (mismatch-forming with respect to target α-1 antitrypsin RNA sequence), mismatch-forming nucleotides can occur consecutively (e.g., in a triplet along the antisense strand nucleotide sequence). Alternatively, nucleotides of the antisense strand that form mismatched base pairs with the target α-1 antitrypsin RNA sequence can be interspersed by nucleotides that form matched base pairs with the target α-1 antitrypsin RNA sequence (e.g., for a DsiRNA possessing mismatched nucleotides at positions 17, 18 and 22, but not at positions 19, 20 and 21, the mismatch-forming residues of antisense strand positions 17 and 18 are adjacent to one another, while the mismatch-forming residues of antisense strand positions 18 and 22 are interspersed by three nucleotides that form matched base pairs with corresponding residues of the target α-1 antitrypsin RNA). For example, three residues of the antisense strand (located within the mismatch-tolerant region of the antisense strand) that form mismatched base pairs with the corresponding target α-1 antitrypsin RNA sequence can occur with zero, one, two, three or four matched base pairs located between any two of these mismatch-forming base pairs.

For certain dsRNAs possessing four mismatch-forming base pairs (mismatch-forming with respect to target α-1 antitrypsin RNA sequence), mismatch-forming nucleotides can occur consecutively (e.g., in a quadruplet along the sense strand nucleotide sequence). Alternatively, nucleotides of the antisense strand that form mismatched base pairs with the target α-1 antitrypsin RNA sequence can be interspersed by nucleotides that form matched base pairs with the target α-1 antitrypsin RNA sequence (e.g., for a DsiRNA possessing mismatch-forming nucleotides at positions 17, 19, 21 and 22, but not at positions 18 and 20, the mismatch-forming residues of antisense strand positions 21 and 22 are adjacent to one another, while the mismatch-forming residues of antisense strand positions 17 and 19 are interspersed by one nucleotide that forms a matched base pair with the corresponding residue of the target α-1 antitrypsin RNA sequence—similarly, the mismatch-forming residues of antisense strand positions 19 and 21 are also interspersed by one nucleotide that forms a matched base pair with the corresponding residue of the target α-1 antitrypsin RNA sequence). For example, four residues of the antisense strand (located within the mismatch-tolerant region of the antisense strand) that form mismatched base pairs with the corresponding target α-1 antitrypsin RNA sequence can occur with zero, one, two or three matched base pairs located between any two of these mismatch-forming base pairs.

The above DsiRNAmm and other dsRNA structures are described in order to exemplify certain structures of DsiRNAmm and dsRNA agents. Design of the above DsiRNAmm and dsRNA structures can be adapted to generate, e.g., DsiRNAmm forms of other DsiRNA structures shown infra. As exemplified above, dsRNAs can also be designed that possess single mismatches (or two, three or four mismatches) between the antisense strand of the dsRNA and a target sequence, yet optionally can retain perfect complementarity between sense and antisense strand sequences of a dsRNA.

It is further noted that the dsRNA agents exemplified infra can also possess insertion/deletion (in/del) structures within their double-stranded and/or target α-1 antitrypsin RNA-aligned structures. Accordingly, the dsRNAs of the invention can be designed to possess in/del variations in, e.g., antisense strand sequence as compared to target α-1 antitrypsin RNA sequence and/or antisense strand sequence as compared to sense strand sequence, with preferred location(s) for placement of such in/del nucleotides corresponding to those locations described above for positioning of mismatched and/or mismatch-forming base pairs.

It is also noted that the DsiRNAs of the instant invention can tolerate mismatches within the 3′-terminal region of the sense strand/5′-terminal region of the antisense strand, as this region is modeled to be processed by Dicer and liberated from the guide strand sequence that loads into RISC. Exemplary DsiRNA structures of the invention that harbor such mismatches include the following:

Target RNA Sequence: 5′. . . XXXXXXXXXXXXXXXXXXXXXHXXX . . .-3′ DsiRNA Sense Strand:      5′-XXXXXXXXXXXXXXXXXXXXXIXDD-3′ DsiRNA Antisense Strand:    3′-XXXXXXXXXXXXXXXXXXXXXXXJXXX-5′ Target RNA Sequence: 5′. . . XXXXXXXXXXXXXXXXXXXXXXHXX . . .-3′ DsiRNA Sense Strand:      5′-XXXXXXXXXXXXXXXXXXXXXXIDD-3′ DsiRNA Antisense Strand:    3′-XXXXXXXXXXXXXXXXXXXXXXXXJXX-5′ Target RNA Sequence: 5′-. . . XXXXXXXXXXXXXXXXXXXXXXXHX . . .-3′ DsiRNA Sense Strand:       5′-XXXXXXXXXXXXXXXXXXXXXXXID-3′ DsiRNA Antisense Strand:     3′-XXXXXXXXXXXXXXXXXXXXXXXXXJX-5′ Target RNA Sequence: 5′. . . XXXXXXXXXXXXXXXXXXXXXXXXH . . .-3′ DsiRNA Sense Strand:      5′-XXXXXXXXXXXXXXXXXXXXXXXDI-3′ DsiRNA Antisense Strand:    3′-XXXXXXXXXXXXXXXXXXXXXXXXXXJ-5′ wherein “X”=RNA, “D”=DNA and “I” and “J”=Nucleic acid residues (RNA, DNA or non-natural or modified nucleic acids) that do not base pair (hydrogen bond) with one another, yet optionally “J” is complementary to target RNA sequence nucleotide “H”. Any of the residues of such agents can optionally be 2′-O-methyl RNA monomers—alternating positioning of 2′-O-methyl RNA monomers that commences from the 3′-terminal residue of the bottom (second) strand, as shown above—or any of the above-described methylation patterns—can also be used in the above DsiRNA agents. The above mismatches can also be combined within the DsiRNAs of the instant invention.

In the below exemplary structures, such mismatches are introduced within the asymmetric AAT-506 DsiRNA (newly-introduced mismatch residues are italicized): AAT-506 25/27mer DsiRNA, mismatch position=19 of sense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAACAUCgc-3′ (SEQ ID NO: 3326) 3′-AUAGAAGAAGAGGGGUCACU0GUAGCG-5′ (SEQ ID NO: 213) Optionally, the mismatched ‘A’ residue of position 19 of the sense strand is alternatively ‘U’ or ‘C’. AAT-506 25/27mer DsiRNA, mismatch position=20 of sense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAG1AUCgc-3′ (SEQ ID NO: 3327) 3′-AUAGAAGAAGAGGGGUCACUCGUAGCG-5′ (SEQ ID NO: 213) Optionally, the mismatched ‘U’ residue of position 20 of the sense strand is alternatively ‘A’ or ‘G’. AAT-506 25/27mer DsiRNA, mismatch position=21 of sense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAGCUUCgc-3′ (SEQ ID NO: 3328) 3′-AUAGAAGAAGAGGGGUCACUCGuAGCG-5′ (SEQ ID NO: 213) Optionally, the mismatched ‘U’ residue of position 21 of the sense strand is alternatively ‘G’ or ‘C’. AAT-506 25/27mer DsiRNA, mismatch position=22 of sense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAGCAGCgc-3′ (SEQ ID NO: 3329) 3′-AUAGAAGAAGAGGGGUCACUCGUAGCG-5′ (SEQ ID NO: 213) Optionally, the mismatched ‘G’ residue of position 22 of the sense strand is alternatively ‘A’ or ‘C’. AAT-506 25/27mer DsiRNA, mismatch position=23 of sense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAGCAUAgc-3′ (SEQ ID NO: 3330) 3′-AUAGAAGAAGAGGGGUCACUCGUAGCG-5′ (SEQ ID NO: 213) Optionally, the mismatched ‘A’ residue of position 23 of the sense strand is alternatively ‘U’ or ‘G’. AAT-506 25/27mer DsiRNA, mismatch position=24 of sense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAGCAUCtc-3′ (SEQ ID NO: 3331) 3′-AUAGAAGAAGAGGGGUCACUCGUAG0G-5′ (SEQ ID NO: 213) Optionally, the mismatched ‘t’ residue of position 24 of the sense strand is alternatively ‘a’ or ‘c’. AAT-506 25/27mer DsiRNA, mismatch position=25 of sense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAGCAUCga-3′ (SEQ ID NO: 3332) 3′-AUAGAAGAAGAGGGGUCACUCGUAGCG-5′ (SEQ ID NO: 213) Optionally, the mismatched ‘a’ residue of position 25 of the sense strand is alternatively ‘t’ or ‘g’. AAT-506 25/27mer DsiRNA, mismatch position=1 of antisense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAGCAUCgc-3′ (SEQ ID NO: 15) 3′-AUAGAAGAAGAGGGGUCACUCGUAGCu-5′ (SEQ ID NO: 3333) Optionally, the mismatched ‘U’ residue of position 1 of the antisense strand is alternatively ‘A’ or ‘C’. AAT-506 25/27mer DsiRNA, mismatch position=2 of antisense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAGCAUCgc-3′ (SEQ ID NO: 15) 3′-AUAGAAGAAGAGGGGUCACUCGUAGAG-5′ (SEQ ID NO: 3334) Optionally, the mismatched ‘A’ residue of position 2 of the antisense strand is alternatively ‘U’ or ‘G’. AAT-506 25/27mer DsiRNA, mismatch position=3 of antisense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAGCAUCgc-3′ (SEQ ID NO: 15) 3′-AUAGAAGAAGAGGGGUCACUCGUAuCG-5′ (SEQ ID NO: 3335) Optionally, the mismatched ‘U’ residue of position 3 of the antisense strand is alternatively ‘A’ or ‘C’. AAT-506 25/27mer DsiRNA, mismatch position=4 of antisense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAGCAUCgc-3′ (SEQ ID NO: 15) 3′-AUAGAAGAAGAGGGGUCACUCGUcGCG-5′ (SEQ ID NO: 3336) Optionally, the mismatched ‘C’ residue of position 4 of the antisense strand is alternatively ‘U’ or ‘G’. AAT-506 25/27mer DsiRNA, mismatch position=5 of antisense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAGCAUCgc-3′ (SEQ ID NO: 15) 3′-AUAGAAGAAGAGGGGUCACUCGAAGCG-5′ (SEQ ID NO: 3337) Optionally, the mismatched ‘A’ residue of position 5 of the antisense strand is alternatively ‘C’ or ‘G’. AAT-506 25/27mer DsiRNA, mismatch position=6 of antisense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAGCAUCgc-3′ (SEQ ID NO: 15) 3′-AUAGAAGAAGAGGGGUCACUCAUAGCG-5′ (SEQ ID NO: 3338) Optionally, the mismatched ‘A’ residue of position 6 of the antisense strand is alternatively ‘U’ or ‘C’. AAT-506 25/27mer DsiRNA, mismatch position=7 of antisense strand (from 5′-terminus)

  5′-UCUUCUUCUCCCCAGUGAGCAUCgc-3′ (SEQ ID NO: 15) 3′-AUAGAAGAAGAGGGGUCACUuGUAGCG-5′ (SEQ ID NO: 3339) Optionally, the mismatched ‘U’ residue of position 7 of the antisense strand is alternatively ‘A’ or ‘G’.

As another example, in the below structures, such mismatches are introduced within the asymmetric AAT-1059 DsiRNA (newly-introduced mismatch residues are italicized): AAT-1059 25/27mer DsiRNA, mismatch position=19 of sense strand (from 5′-terminus)

(SEQ ID NO: 3340) 5′-GCUGUCCAGCUGGGUGCUACUGAtg-3′ (SEQ ID NO: 285) 3′-UUCGACAGGUCGA000ACGA0GACUAC-5′ Optionally, the mismatched ‘A’ residue of position 19 of the sense strand is alternatively ‘U’ or ‘C’. AAT-1059 25/27mer DsiRNA, mismatch position=20 of sense strand (from 5′-terminus)

(SEQ ID NO: 3341) 5′-GCUGUCCAGCUGGGUGCUGUUGAtg-3′ (SEQ ID NO: 285) 3′-UUCGACAGGUCGA000ACGAC0ACUAC-5′ Optionally, the mismatched ‘U’ residue of position 20 of the sense strand is alternatively ‘A’ or ‘G’. AAT-1059 25/27mer DsiRNA, mismatch position=21 of sense strand (from 5′-terminus)

(SEQ ID NO: 3342) 5′-GCUGUCCAGCUGGGUGCUGCAGAtg-3′ (SEQ ID NO: 285) 3′-UUCGACAGGUCGA000ACGACGACUAC-5′ Optionally, the mismatched ‘A’ residue of position 21 of the sense strand is alternatively ‘C’ or ‘G’. AAT-1059 25/27mer DsiRNA, mismatch position=22 of sense strand (from 5′-terminus)

(SEQ ID NO: 3343) 5′-GCUGUCCAGCUGGGUGCUGCUUAtg-3′ (SEQ ID NO: 285) 3′-UUCGACAGGUCGA000ACGACGA0UAC-5′ Optionally, the mismatched ‘U’ residue of position 22 of the sense strand is alternatively ‘A’ or ‘C’. AAT-1059 25/27mer DsiRNA, mismatch position=23 of sense strand (from 5′-terminus)

(SEQ ID NO: 3344) 5′-GCUGUCCAGCUGGGUGCUGCUGUtg-3′ (SEQ ID NO: 285) 3′-UUCGACAGGUCGACCCACGACGACuAC-5′ Optionally, the mismatched ‘U’ residue of position 23 of the sense strand is alternatively ‘C’ or ‘G’. AAT-1059 25/27mer DsiRNA, mismatch position=24 of sense strand (from 5′-terminus)

(SEQ ID NO: 3345) 5′-GCUGUCCAGCUGGGUGCUGCUGAgg-3′ (SEQ ID NO: 285) 3′-UUCGACAGGUCGA000ACGACGACUAC-5′ Optionally, the mismatched ‘g’ residue of position 24 of the sense strand is alternatively ‘a’ or ‘c’. AAT-1059 25/27mer DsiRNA, mismatch position=25 of sense strand (from 5′-terminus)

(SEQ ID NO: 3346) 5′-GCUGUCCAGCUGGGUGCUGCUGAta-3′ (SEQ ID NO: 285) 3′-UUCGACAGGUCGA000ACGACGACUA0-5′ Optionally, the mismatched ‘a’ residue of position 25 of the sense strand is alternatively ‘t’ or ‘c’. AAT-1059 25/27mer DsiRNA, mismatch position=1 of antisense strand (from 5′-terminus)

(SEQ ID NO: 87) 5′-GCUGUCCAGCUGGGUGCUGCUGAtg-3′ (SEQ ID NO: 3347) 3′-UUCGACAGGUCGACCCACGACGACUA 5′ Optionally, the mismatched ‘U’ residue of position 1 of the antisense strand is alternatively ‘A’ or ‘G’. AAT-1059 25/27mer DsiRNA, mismatch position=2 of antisense strand (from 5′-terminus)

(SEQ ID NO: 87) 5′-GCUGUCCAGCUGGGUGCUGCUGAtg-3′ (SEQ ID NO: 3348) 3′-UUCGACAGGUCGA000ACGACGACU0C-5′ Optionally, the mismatched ‘C’ residue of position 2 of the antisense strand is alternatively ‘U’ or ‘G’. AAT-1059 25/27mer DsiRNA, mismatch position=3 of antisense strand (from 5′-terminus)

(SEQ ID NO: 87) 5′-GCUGUCCAGCUGGGUGCUGCUGAtg-3′ (SEQ ID NO: 3349) 3′-UUCGACAGGUCGA000ACGACGACAAC-5′ Optionally, the mismatched ‘A’ residue of position 3 of the antisense strand is alternatively ‘C’ or ‘G’. AAT-1059 25/27mer DsiRNA, mismatch position=4 of antisense strand (from 5′-terminus)

(SEQ ID NO: 87) 5′-GCUGUCCAGCUGGGUGCUGCUGAtg-3′ (SEQ ID NO: 3350) 3′-UUCGACAGGUCGA000ACGACGAAUAC-5′ Optionally, the mismatched ‘A’ residue of position 4 of the antisense strand is alternatively ‘U’ or ‘G’. AAT-1059 25/27mer DsiRNA, mismatch position=5 of antisense strand (from 5′-terminus)

(SEQ ID NO: 87) 5′-GCUGUCCAGCUGGGUGCUGCUGAtg-3′ (SEQ ID NO: 3351) 3′-UUCGACAGGUCGACCCACGACGuCUAC-5′ Optionally, the mismatched ‘U’ residue of position 5 of the antisense strand is alternatively ‘C’ or ‘G’. AAT-1059 25/27mer DsiRNA, mismatch position=6 of antisense strand (from 5′-terminus)

(SEQ ID NO: 87) 5′-GCUGUCCAGCUGGGUGCUGCUGAtg-3′ (SEQ ID NO: 3352) 3′-UUCGACAGGUCGA000ACGACAACUAC-5′ Optionally, the mismatched ‘A’ residue of position 6 of the antisense strand is alternatively ‘U’ or ‘C’. AAT-1059 25/27mer DsiRNA, mismatch position=7 of antisense strand (from 5′-terminus)

(SEQ ID NO: 87) 5′-GCUGUCCAGCUGGGUGCUGCUGAtg-3′ (SEQ ID NO: 3353) 3′-UUCGACAGGUCGACCCACGAuGACUAC-5′ Optionally, the mismatched ‘U’ residue of position 7 of the antisense strand is alternatively ‘A’ or ‘G’.

For the above oligonucleotide strand sequences, it is contemplated that the sense strand sequence of one depicted duplex can be combined with an antisense strand of another depicted duplex, thereby forming a distinct duplex—in certain instances, such duplexes contain a mismatched residue with respect to the α-1 antitrypsin target transcript sequence, while such sense and antisense strand sequences do not present a mismatch at this residue with respect to one another (e.g., duplexes comprising SEQ ID NOs: 3326 and 3339; SEQ ID NOs: 3327 and 3338; SEQ ID NOs: 3328 and 3337, etc., are contemplated as exemplary of such duplexes).

As noted above, introduction of mismatches can be performed upon any of the DsiRNAs described herein.

The mismatches of such DsiRNA structures can be combined to produce a DsiRNA possessing, e.g., two, three or even four mismatches within the 3′-terminal four to seven nucleotides of the sense strand/5′-terminal four to seven nucleotides of the antisense strand.

Indeed, in view of the flexibility of sequences which can be incorporated into DsiRNAs at the 3′-terminal residues of the sense strand/5′-terminal residues of the antisense strand, in certain embodiments, the sequence requirements of an asymmetric DsiRNA of the instant invention can be represented as the following (minimalist) structure (shown for an exemplary α-1 antitrypsin-506 DsiRNA sequence):

(SEQ ID NO: 3354) 5′-UCUUCUUCUCCCCAGUGAXXXXXX[X]n-3′ (SEQ ID NO: 3355) 3′-AUAGAAGAAGAGGGGUCACUXXXXXX[X],-5′ where n=1 to 5, 1 to 10, 1 to 20, 1 to 30, 1 to 50, or 1 to 80 or more. α-1 antitrypsin-506 mRNA Target: 5′-UAUCUUCUUCUCCCCAGUGAXXXXXXX-3′ (SEQ ID NO: 3356).

The α-1 antitrypsin target site may also be a site which is targeted by one or more of several oligonucleotides whose complementary target sites overlap with a stated target site. For example, for an exemplary α-1 antitrypsin-506 DsiRNA, it is noted that certain DsiRNAs targeting overlapping and only slightly offset α-1 antitrypsin sequences could exhibit activity levels similar to that of α-1 antitrypsin-506 (e.g., α-1 antitrypsin-500 to 513 of Table 2 above). Thus, in certain embodiments, a designated target sequence region might be effectively targeted by a series of DsiRNAs possessing largely overlapping sequences. (E.g., if considering DsiRNAs of the α-1 antitrypsin-500 to α-1 antitrypsin-513 target site(s), a more encompassing α-1 antitrypsin transcript target sequence might be recited as, e.g., 5′-CACCAAUAUCUUCUUCUCCCCAGUGAGCAUCGCU-3′ (SEQ ID NO: 3357), wherein any given DsiRNA (e.g., a DsiRNA selected from α-1 antitrypsin-500 to α-1 antitrypsin-513) only targets a sub-sequence within such a sequence region, yet the entire sequence can be considered a viable target for such a series of DsiRNAs).

Additionally and/or alternatively, mismatches within the 3′-terminal seven nucleotides of the sense strand/5′-terminal seven nucleotides of the antisense strand can be combined with mismatches positioned at other mismatch-tolerant positions, as described above.

In view of the present identification of the above-described Dicer substrate agents (DsiRNAs) as inhibitors of α-1 antitrypsin levels via targeting of specific α-1 antitrypsin sequences, it is also recognized that dsRNAs having structures similar to those described herein can also be synthesized which target other sequences within the α-1 antitrypsin sequence of NM_000295.4, or within variants thereof (e.g., target sequences possessing 80% identity, 90% identity, 95% identity, 96% identity, 97% identity, 98% identity, 99% or more identity to a sequence of NM_000295.4).

Anti-α-1 Antitrypsin DsiRNA Design/Synthesis

It has been found empirically that longer dsRNA species of from 25 to 35 nucleotides (DsiRNAs) and especially from 25 to 30 nucleotides give unexpectedly effective results in terms of potency and duration of action, as compared to 19-23mer siRNA agents. Without wishing to be bound by the underlying theory of the dsRNA processing mechanism, it is thought that the longer dsRNA species serve as a substrate for the Dicer enzyme in the cytoplasm of a cell. In addition to cleaving the dsRNA of the invention into shorter segments, Dicer is thought to facilitate the incorporation of a single-stranded cleavage product derived from the cleaved dsRNA into the RISC complex that is responsible for the destruction of the cytoplasmic RNA (e.g., α-1 antitrypsin RNA) of or derived from the target gene, α-1 antitrypsin (or other gene associated with a α-1 antitrypsin-associated disease or disorder). Prior studies (Rossi et al., U.S. Patent Application No. 2007/0265220) have shown that the cleavability of a dsRNA species (specifically, a DsiRNA agent) by Dicer corresponds with increased potency and duration of action of the dsRNA species.

Certain preferred anti-α-1 antitrypsin DsiRNA agents were selected from a pre-screened population. Design of DsiRNAs can optionally involve use of predictive scoring algorithms that perform in silico assessments of the projected activity/efficacy of a number of possible DsiRNA agents spanning a region of sequence. Information regarding the design of such scoring algorithms can be found, e.g., in Gong et al. (BMC Bioinformatics 2006, 7:516), though a more recent “v4.3” algorithm represents a theoretically improved algorithm relative to siRNA scoring algorithms previously available in the art. (E.g., “v3” and “v4” scoring algorithms are machine learning algorithms that are not reliant upon any biases in human sequence. In addition, the “v3” and “v4” algorithms derive from data sets that are many-fold larger than that from which an older “v2” algorithm such as that described in Gong et al. derives.)

The first and second oligonucleotides of the DsiRNA agents of the instant invention are not required to be completely complementary. In fact, in one embodiment, the 3′-terminus of the sense strand contains one or more mismatches. In one aspect, two mismatches are incorporated at the 3′ terminus of the sense strand. In another embodiment, the DsiRNA of the invention is a double stranded RNA molecule containing two RNA oligonucleotides each of which is 27 nucleotides in length and, when annealed to each other, have blunt ends and a two nucleotide mismatch on the 3′-terminus of the sense strand (the 5′-terminus of the antisense strand). The use of mismatches or decreased thermodynamic stability (specifically at the 3′-sense/5′-antisense position) has been proposed to facilitate or favor entry of the antisense strand into RISC (Schwarz et al., 2003, Cell 115: 199-208; Khvorova et al., 2003, Cell 115: 209-216), presumably by affecting some rate-limiting unwinding steps that occur with entry of the siRNA into RISC. Thus, terminal base composition has been included in design algorithms for selecting active 21mer siRNA duplexes (Ui-Tei et al., 2004, Nucleic Acids Res 32: 936-948; Reynolds et al., 2004, Nat Biotechnol 22: 326-330). With Dicer cleavage of the dsRNA of this embodiment, the small end-terminal sequence which contains the mismatches will either be left unpaired with the antisense strand (become part of a 3′-overhang) or be cleaved entirely off the final 21-mer siRNA. These “mismatches”, therefore, do not persist as mismatches in the final RNA component of RISC. The finding that base mismatches or destabilization of segments at the 3′-end of the sense strand of Dicer substrate improved the potency of synthetic duplexes in RNAi, presumably by facilitating processing by Dicer, was a surprising finding of past works describing the design and use of 25-30mer dsRNAs (also termed “DsiRNAs” herein; Rossi et al., U.S. Patent Application Nos. 2005/0277610, 2005/0244858 and 2007/0265220).

Modification of Anti-α-1 Antitrypsin dsRNAs

One major factor that inhibits the effect of double stranded RNAs (“dsRNAs”) is the degradation of dsRNAs (e.g., siRNAs and DsiRNAs) by nucleases. A 3′-exonuclease is the primary nuclease activity present in serum and modification of the 3′-ends of antisense DNA oligonucleotides is crucial to prevent degradation (Eder et al., 1991, Antisense Res Dev, 1: 141-151). An RNase-T family nuclease has been identified called ERI-1 which has 3′ to 5′ exonuclease activity that is involved in regulation and degradation of siRNAs (Kennedy et al., 2004, Nature 427: 645-649; Hong et al., 2005, Biochem J, 390: 675-679). This gene is also known as Thex1 (NM_02067) in mice or THEX1 (NM_153332) in humans and is involved in degradation of histone mRNA; it also mediates degradation of 3′-overhangs in siRNAs, but does not degrade duplex RNA (Yang et al., 2006, J Biol Chem, 281: 30447-30454). It is therefore reasonable to expect that 3′-end-stabilization of dsRNAs, including the DsiRNAs of the instant invention, will improve stability.

XRN1 (NM_019001) is a 5′ to 3′ exonuclease that resides in P-bodies and has been implicated in degradation of mRNA targeted by miRNA (Rehwinkel et al., 2005, RNA 11: 1640-1647) and may also be responsible for completing degradation initiated by internal cleavage as directed by a siRNA. XRN2 (NM_012255) is a distinct 5′ to 3′ exonuclease that is involved in nuclear RNA processing.

RNase A is a major endonuclease activity in mammals that degrades RNAs. It is specific for ssRNA and cleaves at the 3′-end of pyrimidine bases. SiRNA degradation products consistent with RNase A cleavage can be detected by mass spectrometry after incubation in serum (Turner et al., 2007, Mol Biosyst 3: 43-50). The 3′-overhangs enhance the susceptibility of siRNAs to RNase degradation. Depletion of RNase A from serum reduces degradation of siRNAs; this degradation does show some sequence preference and is worse for sequences having poly A/U sequence on the ends (Haupenthal et al., 2006 Biochem Pharmacol 71: 702-710). This suggests the possibility that lower stability regions of the duplex may “breathe” and offer transient single-stranded species available for degradation by RNase A. RNase A inhibitors can be added to serum and improve siRNA longevity and potency (Haupenthal et al., 2007, Int J. Cancer 121: 206-210).

In 21mers, phosphorothioate or boranophosphate modifications directly stabilize the internucleoside phosphate linkage. Boranophosphate modified RNAs are highly nuclease resistant, potent as silencing agents, and are relatively non-toxic. Boranophosphate modified RNAs cannot be manufactured using standard chemical synthesis methods and instead are made by in vitro transcription (IVT) (Hall et al., 2004, Nucleic Acids Res 32: 5991-6000; Hall et al., 2006, Nucleic Acids Res 34: 2773-2781). Phosphorothioate (PS) modifications can be easily placed in the RNA duplex at any desired position and can be made using standard chemical synthesis methods. The PS modification shows dose-dependent toxicity, so most investigators have recommended limited incorporation in siRNAs, favoring the 3′-ends where protection from nucleases is most important (Harborth et al., 2003, Antisense Nucleic Acid Drug Dev 13: 83-105; Chiu and Rana, 2003, Mol Cell 10: 549-561; Braasch et al., 2003, Biochemistry 42: 7967-7975; Amarzguioui et al., 2003, Nucleic Acids Research 31: 589-595). More extensive PS modification can be compatible with potent RNAi activity; however, use of sugar modifications (such as 2′-O-methyl RNA) may be superior (Choung et al., 2006, Biochem Biophys Res Commun 342: 919-927).

A variety of substitutions can be placed at the 2′-position of the ribose which generally increases duplex stability (T_(m)) and can greatly improve nuclease resistance. 2′-O-methyl RNA is a naturally occurring modification found in mammalian ribosomal RNAs and transfer RNAs. 2′-O-methyl modification in siRNAs is known, but the precise position of modified bases within the duplex is important to retain potency and complete substitution of 2′-O-methyl RNA for RNA will inactivate the siRNA. For example, a pattern that employs alternating 2′-O-methyl bases can have potency equivalent to unmodified RNA and is quite stable in serum (Choung et al., 2006, Biochem Biophys Res Commun 342: 919-927; Czauderna et al., 2003, Nucleic Acids Research 31: 2705-2716).

The 2′-fluoro (2′-F) modification is also compatible with dsRNA (e.g., siRNA and DsiRNA) function; it is most commonly placed at pyrimidine sites (due to reagent cost and availability) and can be combined with 2′-O-methyl modification at purine positions; 2′-F purines are available and can also be used. Heavily modified duplexes of this kind can be potent triggers of RNAi in vitro (Allerson et al., 2005, J Med Chem 48: 901-904; Prakash et al., 2005, J Med Chem 48: 4247-4253; Kraynack and Baker, 2006, RNA 12: 163-176) and can improve performance and extend duration of action when used in vivo (Morrissey et al., 2005, Hepatology 41: 1349-1356; Morrissey et al., 2005, Nat Biotechnol 23: 1002-1007). A highly potent, nuclease stable, blunt 19mer duplex containing alternative 2′-F and 2′-O-Me bases is taught by Allerson. In this design, alternating 2′-O-Me residues are positioned in an identical pattern to that employed by Czauderna, however the remaining RNA residues are converted to 2′-F modified bases. A highly potent, nuclease resistant siRNA employed by Morrissey employed a highly potent, nuclease resistant siRNA in vivo. In addition to 2′-O-Me RNA and 2′-F RNA, this duplex includes DNA, RNA, inverted abasic residues, and a 3′-terminal PS internucleoside linkage. While extensive modification has certain benefits, more limited modification of the duplex can also improve in vivo performance and is both simpler and less costly to manufacture. Soutschek et al. (2004, Nature 432: 173-178) employed a duplex in vivo and was mostly RNA with two 2′-O-Me RNA bases and limited 3′-terminal PS internucleoside linkages.

Locked nucleic acids (LNAs) are a different class of 2′-modification that can be used to stabilize dsRNA (e.g., siRNA and DsiRNA). Patterns of LNA incorporation that retain potency are more restricted than 2′-O-methyl or 2′-F bases, so limited modification is preferred (Braasch et al., 2003, Biochemistry 42: 7967-7975; Grunweller et al., 2003, Nucleic Acids Res 31: 3185-3193; Elmen et al., 2005, Nucleic Acids Res 33: 439-447). Even with limited incorporation, the use of LNA modifications can improve dsRNA performance in vivo and may also alter or improve off target effect profiles (Mook et al., 2007, Mol Cancer Ther 6: 833-843).

Synthetic nucleic acids introduced into cells or live animals can be recognized as “foreign” and trigger an immune response. Immune stimulation constitutes a major class of off-target effects which can dramatically change experimental results and even lead to cell death. The innate immune system includes a collection of receptor molecules that specifically interact with DNA and RNA that mediate these responses, some of which are located in the cytoplasm and some of which reside in endosomes (Marques and Williams, 2005, Nat Biotechnol 23: 1399-1405; Schlee et al., 2006, Mol Ther 14: 463-470). Delivery of siRNAs by cationic lipids or liposomes exposes the siRNA to both cytoplasmic and endosomal compartments, maximizing the risk for triggering a type 1 interferon (IFN) response both in vitro and in vivo (Morrissey et al., 2005, Nat Biotechnol 23: 1002-1007; Sioud and Sorensen, 2003, Biochem Biophys Res Commun 312: 1220-1225; Sioud, 2005, J Mol Biol 348: 1079-1090; Ma et al., 2005, Biochem Biophys Res Commun 330: 755-759). RNAs transcribed within the cell are less immunogenic (Robbins et al., 2006, Nat Biotechnol 24: 566-571) and synthetic RNAs that are immunogenic when delivered using lipid-based methods can evade immune stimulation when introduced unto cells by mechanical means, even in vivo (Heidel et al., 2004, Nat Biotechnol 22: 1579-1582). However, lipid based delivery methods are convenient, effective, and widely used. Some general strategy to prevent immune responses is needed, especially for in vivo application where all cell types are present and the risk of generating an immune response is highest. Use of chemically modified RNAs may solve most or even all of these problems.

In certain embodiments, modifications can be included in the anti-α-1 antitrypsin dsRNA agents of the present invention so long as the modification does not prevent the dsRNA agent from possessing α-1 antitrypsin inhibitory activity. In one embodiment, one or more modifications are made that enhance Dicer processing of the DsiRNA agent (an assay for determining Dicer processing of a DsiRNA is described elsewhere herein). In a second embodiment, one or more modifications are made that result in more effective α-1 antitrypsin inhibition (as described herein, α-1 antitrypsin inhibition/α-1 antitrypsin inhibitory activity of a dsRNA can be assayed via art-recognized methods for determining RNA levels, or for determining α-1 antitrypsin polypeptide levels, should such levels be assessed in lieu of or in addition to assessment of, e.g., α-1 antitrypsin mRNA levels). In a third embodiment, one or more modifications are made that support greater α-1 antitrypsin inhibitory activity (means of determining α-1 antitrypsin inhibitory activity are described supra). In a fourth embodiment, one or more modifications are made that result in greater potency of α-1 antitrypsin inhibitory activity per each dsRNA agent molecule to be delivered to the cell (potency of α-1 antitrypsin inhibitory activity is described supra). Modifications can be incorporated in the 3′-terminal region, the 5′-terminal region, in both the 3′-terminal and 5′-terminal region or in some instances in various positions within the sequence. With the restrictions noted above in mind, numbers and combinations of modifications can be incorporated into the dsRNA agent. Where multiple modifications are present, they may be the same or different. Modifications to bases, sugar moieties, the phosphate backbone, and their combinations are contemplated. Either 5′-terminus can be phosphorylated.

Examples of modifications contemplated for the phosphate backbone include phosphonates, including methylphosphonate, phosphorothioate, and phosphotriester modifications such as alkylphosphotriesters, and the like. Examples of modifications contemplated for the sugar moiety include 2′-alkyl pyrimidine, such as 2′-O-methyl, 2′-fluoro, amino, and deoxy modifications and the like (see, e.g., Amarzguioui et al., 2003, Nucleic Acids Research 31: 589-595). Examples of modifications contemplated for the base groups include abasic sugars, 2-O-alkyl modified pyrimidines, 4-thiouracil, 5-bromouracil, 5-iodouracil, and 5-(3-aminoallyl)-uracil and the like. Locked nucleic acids, or LNA's, could also be incorporated. Many other modifications are known and can be used so long as the above criteria are satisfied. Examples of modifications are also disclosed in U.S. Pat. Nos. 5,684,143, 5,858,988 and 6,291,438 and in U.S. published patent application No. 2004/0203145 A1. Other modifications are disclosed in Herdewijn (2000, Antisense Nucleic Acid Drug Dev 10: 297-310), Eckstein (2000, Antisense Nucleic Acid Drug Dev 10: 117-21), Rusckowski et al. (2000, Antisense Nucleic Acid Drug Dev 10: 333-345), Stein et al. (2001, Antisense Nucleic Acid Drug Dev 11: 317-25); Vorobjev et al. (2001, Antisense Nucleic Acid Drug Dev 11: 77-85).

One or more modifications contemplated can be incorporated into either strand. The placement of the modifications in the dsRNA agent can greatly affect the characteristics of the dsRNA agent, including conferring greater potency and stability, reducing toxicity, enhance Dicer processing, and minimizing an immune response. In one embodiment, the antisense strand or the sense strand or both strands have one or more 2′-O-methyl modified nucleotides. In another embodiment, the antisense strand contains 2′-O-methyl modified nucleotides. In another embodiment, the antisense stand contains a 3′ overhang that is comprised of 2′-O-methyl modified nucleotides. The antisense strand could also include additional 2′-O-methyl modified nucleotides.

In certain embodiments, the anti-α-1 antitrypsin DsiRNA agent of the invention has several properties which enhance its processing by Dicer. According to such embodiments, the DsiRNA agent has a length sufficient such that it is processed by Dicer to produce an siRNA and at least one of the following properties: (i) the DsiRNA agent is asymmetric, e.g., has a 3′ overhang on the sense strand and (ii) the DsiRNA agent has a modified 3′ end on the antisense strand to direct orientation of Dicer binding and processing of the dsRNA to an active siRNA. According to these embodiments, the longest strand in the DsiRNA agent comprises 25-30 nucleotides. In one embodiment, the sense strand comprises 25-30 nucleotides and the antisense strand comprises 25-28 nucleotides. Thus, the resulting dsRNA has an overhang on the 3′ end of the sense strand. The overhang is 1˜4 nucleotides, such as 2 nucleotides. The antisense strand may also have a 5′ phosphate.

In certain embodiments, the sense strand of a DsiRNA agent is modified for Dicer processing by suitable modifiers located at the 3′ end of the sense strand, i.e., the DsiRNA agent is designed to direct orientation of Dicer binding and processing. Suitable modifiers include nucleotides such as deoxyribonucleotides, dideoxyribonucleotides, acyclonucleotides and the like and sterically hindered molecules, such as fluorescent molecules and the like. Acyclonucleotides substitute a 2-hydroxyethoxymethyl group for the 2′-deoxyribofuranosyl sugar normally present in dNMPs. Other nucleotide modifiers could include 3′-deoxyadenosine (cordycepin), 3′-azido-3′-deoxythymidine (AZT), 2′,3′-dideoxyinosine (ddI), 2′,3′-dideoxy-3′-thiacytidine (3TC), 2′,3′-didehydro-2′,3′-dideoxythymidine (d4T) and the monophosphate nucleotides of 3′-azido-3′-deoxythymidine (AZT), 2′,3′-dideoxy-3′-thiacytidine (3TC) and 2′,3′-didehydro-2′,3′-dideoxythymidine (d4T). In one embodiment, deoxynucleotides are used as the modifiers. When nucleotide modifiers are utilized, 1-3 nucleotide modifiers, or 2 nucleotide modifiers are substituted for the ribonucleotides on the 3′ end of the sense strand. When sterically hindered molecules are utilized, they are attached to the ribonucleotide at the 3′ end of the antisense strand. Thus, the length of the strand does not change with the incorporation of the modifiers. In another embodiment, the invention contemplates substituting two DNA bases in the dsRNA to direct the orientation of Dicer processing. In a further invention, two terminal DNA bases are located on the 3′ end of the sense strand in place of two ribonucleotides forming a blunt end of the duplex on the 5′ end of the antisense strand and the 3′ end of the sense strand, and a two-nucleotide RNA overhang is located on the 3′-end of the antisense strand. This is an asymmetric composition with DNA on the blunt end and RNA bases on the overhanging end.

In certain other embodiments, the antisense strand of a DsiRNA agent is modified for Dicer processing by suitable modifiers located at the 3′ end of the antisense strand, i.e., the DsiRNA agent is designed to direct orientation of Dicer binding and processing. Suitable modifiers include nucleotides such as deoxyribonucleotides, dideoxyribonucleotides, acyclonucleotides and the like and sterically hindered molecules, such as fluorescent molecules and the like. Acyclonucleotides substitute a 2-hydroxyethoxymethyl group for the 2′-deoxyribofuranosyl sugar normally present in dNMPs. Other nucleotide modifiers could include 3′-deoxyadenosine (cordycepin), 3′-azido-3′-deoxythymidine (AZT), 2′,3′-dideoxyinosine (ddI), 2′,3′-dideoxy-3′-thiacytidine (3TC), 2′,3′-didehydro-2′,3′-dideoxythymidine (d4T) and the monophosphate nucleotides of 3′-azido-3′-deoxythymidine (AZT), 2′,3′-dideoxy-3′-thiacytidine (3TC) and 2′,3′-didehydro-2′,3′-dideoxythymidine (d4T). In one embodiment, deoxynucleotides are used as the modifiers. When nucleotide modifiers are utilized, 1-3 nucleotide modifiers, or 2 nucleotide modifiers are substituted for the ribonucleotides on the 3′ end of the antisense strand. When sterically hindered molecules are utilized, they are attached to the ribonucleotide at the 3′ end of the antisense strand. Thus, the length of the strand does not change with the incorporation of the modifiers. In another embodiment, the invention contemplates substituting two DNA bases in the dsRNA to direct the orientation of Dicer processing. In a further invention, two terminal DNA bases are located on the 3′ end of the antisense strand in place of two ribonucleotides forming a blunt end of the duplex on the 5′ end of the sense strand and the 3′ end of the antisense strand, and a two-nucleotide RNA overhang is located on the 3′-end of the sense strand. This is also an asymmetric composition with DNA on the blunt end and RNA bases on the overhanging end.

The sense and antisense strands anneal under biological conditions, such as the conditions found in the cytoplasm of a cell. In addition, a region of one of the sequences, particularly of the antisense strand, of the dsRNA has a sequence length of at least 19 nucleotides, wherein these nucleotides are adjacent to the 3′ end of antisense strand and are sufficiently complementary to a nucleotide sequence of the target α-1 antitryp sin RNA.

Additionally, the DsiRNA agent structure can be optimized to ensure that the oligonucleotide segment generated from Dicer's cleavage will be the portion of the oligonucleotide that is most effective in inhibiting gene expression. For example, in one embodiment of the invention, a 27-bp oligonucleotide of the DsiRNA agent structure is synthesized wherein the anticipated 21 to 22-bp segment that will inhibit gene expression is located on the 3′-end of the antisense strand. The remaining bases located on the 5′-end of the antisense strand will be cleaved by Dicer and will be discarded. This cleaved portion can be homologous (i.e., based on the sequence of the target sequence) or non-homologous and added to extend the nucleic acid strand.

US 2007/0265220 discloses that 27mer DsiRNAs showed improved stability in serum over comparable 21mer siRNA compositions, even absent chemical modification. Modifications of DsiRNA agents, such as inclusion of 2′-O-methyl RNA in the antisense strand, in patterns such as detailed above, when coupled with addition of a 5′ Phosphate, can improve stability of DsiRNA agents. Addition of 5′-phosphate to all strands in synthetic RNA duplexes may be an inexpensive and physiological method to confer some limited degree of nuclease stability.

The chemical modification patterns of the dsRNA agents of the instant invention are designed to enhance the efficacy of such agents. Accordingly, such modifications are designed to avoid reducing potency of dsRNA agents; to avoid interfering with Dicer processing of DsiRNA agents; to improve stability in biological fluids (reduce nuclease sensitivity) of dsRNA agents; or to block or evade detection by the innate immune system. Such modifications are also designed to avoid being toxic and to avoid increasing the cost or impact the ease of manufacturing the instant dsRNA agents of the invention.

In certain embodiments of the present invention, an anti-α-1 antitrypsin DsiRNA agent has one or more of the following properties: (i) the DsiRNA agent is asymmetric, e.g., has a 3′ overhang on the antisense strand and (ii) the DsiRNA agent has a modified 3′ end on the sense strand to direct orientation of Dicer binding and processing of the dsRNA to an active siRNA. According to this embodiment, the longest strand in the dsRNA comprises 25-35 nucleotides (e.g., 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35 nucleotides). In certain such embodiments, the DsiRNA agent is asymmetric such that the sense strand comprises 25-34 nucleotides and the 3′ end of the sense strand forms a blunt end with the 5′ end of the antisense strand while the antisense strand comprises 26-35 nucleotides and forms an overhang on the 3′ end of the antisense strand. In one embodiment, the DsiRNA agent is asymmetric such that the sense strand comprises 25-28 nucleotides and the antisense strand comprises 25-30 nucleotides. Thus, the resulting dsRNA has an overhang on the 3′ end of the antisense strand. The overhang is 1˜4 nucleotides, for example 2 nucleotides. The sense strand may also have a 5′ phosphate.

The DsiRNA agent can also have one or more of the following additional properties: (a) the antisense strand has a right shift from the typical 21mer (e.g., the DsiRNA comprises a length of antisense strand nucleotides that extends to the 5′ of a projected Dicer cleavage site within the DsiRNA, with such antisense strand nucleotides base paired with corresponding nucleotides of the sense strand extending 3′ of a projected Dicer cleavage site in the sense strand), (b) the strands may not be completely complementary, i.e., the strands may contain simple mismatched base pairs (in certain embodiments, the DsiRNAs of the invention possess 1, 2, 3, 4 or even 5 or more mismatched base pairs, provided that α-1 antitrypsin inhibitory activity of the DsiRNA possessing mismatched base pairs is retained at sufficient levels (e.g., retains at least 50% α-1 antitrypsin inhibitory activity or more, at least 60% α-1 antitrypsin inhibitory activity or more, at least 70% α-1 antitrypsin inhibitory activity or more, at least 80% α-1 antitrypsin inhibitory activity or more, at least 90% α-1 antitrypsin inhibitory activity or more or at least 95% α-1 antitrypsin inhibitory activity or more as compared to a corresponding DsiRNA not possessing mismatched base pairs. In certain embodiments, mismatched base pairs exist between the antisense and sense strands of a DsiRNA. In some embodiments, mismatched base pairs exist (or are predicted to exist) between the antisense strand and the target RNA. In certain embodiments, the presence of a mismatched base pair(s) between an antisense strand residue and a corresponding residue within the target RNA that is located 3′ in the target RNA sequence of a projected Ago2 cleavage site retains and may even enhance α-1 antitrypsin inhibitory activity of a DsiRNA of the invention) and (c) base modifications such as locked nucleic acid(s) may be included in the 5′ end of the sense strand. A “typical” 21mer siRNA is designed using conventional techniques. In one technique, a variety of sites are commonly tested in parallel or pools containing several distinct siRNA duplexes specific to the same target with the hope that one of the reagents will be effective (Ji et al., 2003, FEBS Lett 552: 247-252). Other techniques use design rules and algorithms to increase the likelihood of obtaining active RNAi effector molecules (Schwarz et al., 2003, Cell 115: 199-208; Khvorova et al., 2003, Cell 115: 209-216; Ui-Tei et al., 2004, Nucleic Acids Res 32: 936-948; Reynolds et al., 2004, Nat Biotechnol 22: 326-330; Krol et al., 2004, J Biol Chem 279: 42230-42239; Yuan et al., 2004, Nucl Acids Res 32 (Webserver issue):W130-134; Boese et al., 2005, Methods Enzymol 392: 73-96). High throughput selection of siRNA has also been developed (U.S. published patent application No. 2005/0042641 A1). Potential target sites can also be analyzed by secondary structure predictions (Heale et al., 2005, Nucleic Acids Res 33(3): e30). This 21mer is then used to design a right shift to include 3-9 additional nucleotides on the 5′ end of the 21mer. The sequence of these additional nucleotides is not restricted. In one embodiment, the added ribonucleotides are based on the sequence of the target gene. Even in this embodiment, full complementarity between the target sequence and the antisense siRNA is not required.

The first and second oligonucleotides of a DsiRNA agent of the instant invention are not required to be completely complementary. They only need to be sufficiently complementary to anneal under biological conditions and to provide a substrate for Dicer that produces a siRNA sufficiently complementary to the target sequence. Locked nucleic acids, or LNA's, are well known to a skilled artisan (Elmen et al., 2005, Nucleic Acids Res 33: 439-447; Kurreck et al., 2002, Nucleic Acids Res 30: 1911-1918; Crinelli et al., 2002, Nucleic Acids Res 30: 2435-2443; Braasch and Corey, 2001, Chem Biol 8: 1-7; Bondensgaard et al., 2000, Chemistry 6: 2687-2695; Wahlestedt et al., 2000, Proc Natl Acad Sci USA 97: 5633-5638). In one embodiment, an LNA is incorporated at the 5′ terminus of the sense strand. In another embodiment, an LNA is incorporated at the 5′ terminus of the sense strand in duplexes designed to include a 3′ overhang on the antisense strand.

In certain embodiments, the DsiRNA agent of the instant invention has an asymmetric structure, with the sense strand having a 25-base pair length, and the antisense strand having a 27-base pair length with a 2 base 3′-overhang. In other embodiments, this DsiRNA agent having an asymmetric structure further contains 2 deoxynucleotides at the 3′ end of the sense strand in place of two of the ribonucleotides.

Certain DsiRNA agent compositions containing two separate oligonucleotides can be linked by a third structure. The third structure will not block Dicer activity on the DsiRNA agent and will not interfere with the directed destruction of the RNA transcribed from the target gene. In one embodiment, the third structure may be a chemical linking group. Many suitable chemical linking groups are known in the art and can be used. Alternatively, the third structure may be an oligonucleotide that links the two oligonucleotides of the DsiRNA agent in a manner such that a hairpin structure is produced upon annealing of the two oligonucleotides making up the dsRNA composition. The hairpin structure will not block Dicer activity on the DsiRNA agent and will not interfere with the directed destruction of the α-1 antitrypsin RNA.

In Vitro Assay to Assess dsRNA α-1 Antitrypsin Inhibitory Activity

An in vitro assay that recapitulates RNAi in a cell-free system can be used to evaluate dsRNA constructs targeting α-1 antitrypsin RNA sequence(s), and thus to assess α-1 antitrypsin-specific gene inhibitory activity (also referred to herein as α-1 antitrypsin inhibitory activity) of a dsRNA. The assay comprises the system described by Tuschl et al., 1999, Genes and Development, 13, 3191-3197 and Zamore et al., 2000, Cell, 101, 25-33 adapted for use with dsRNA (e.g., DsiRNA) agents directed against α-1 antitrypsin RNA. A Drosophila extract derived from syncytial blastoderm is used to reconstitute RNAi activity in vitro. Target RNA is generated via in vitro transcription from a selected α-1 antitrypsin expressing plasmid using T7 RNA polymerase or via chemical synthesis. Sense and antisense dsRNA strands (for example, 20 uM each) are annealed by incubation in buffer (such as 100 mM potassium acetate, 30 mM HEPES-KOH, pH 7.4, 2 mM magnesium acetate) for 1 minute at 90° C. followed by 1 hour at 37° C., then diluted in lysis buffer (for example 100 mM potassium acetate, 30 mM HEPES-KOH at pH 7.4, 2 mM magnesium acetate). Annealing can be monitored by gel electrophoresis on an agarose gel in TBE buffer and stained with ethidium bromide. The Drosophila lysate is prepared using zero to two-hour-old embryos from Oregon R flies collected on yeasted molasses agar that are dechorionated and lysed. The lysate is centrifuged and the supernatant isolated. The assay comprises a reaction mixture containing 50% lysate [vol/vol], RNA (10-50 pM final concentration), and 10% [vol/vol] lysis buffer containing dsRNA (10 nM final concentration). The reaction mixture also contains 10 mM creatine phosphate, 10 ug/ml creatine phosphokinase, 100 um GTP, 100 uM UTP, 100 uM CTP, 500 uM ATP, 5 mM DTT, 0.1 U/uL RNasin (Promega), and 100 uM of each amino acid. The final concentration of potassium acetate is adjusted to 100 mM. The reactions are pre-assembled on ice and preincubated at 25° C. for 10 minutes before adding RNA, then incubated at 25° C. for an additional 60 minutes. Reactions are quenched with 4 volumes of 1.25× Passive Lysis Buffer (Promega). Target RNA cleavage is assayed by RT-PCR analysis or other methods known in the art and are compared to control reactions in which dsRNA is omitted from the reaction.

Alternately, internally-labeled target RNA for the assay is prepared by in vitro transcription in the presence of [α-³²P] CTP, passed over a G50 Sephadex column by spin chromatography and used as target RNA without further purification. Optionally, target RNA is 5′-³²P-end labeled using T4 polynucleotide kinase enzyme. Assays are performed as described above and target RNA and the specific RNA cleavage products generated by RNAi are visualized on an autoradiograph of a gel. The percentage of cleavage is determined by PHOSPHOR IMAGER® (autoradiography) quantitation of bands representing intact control RNA or RNA from control reactions without dsRNA and the cleavage products generated by the assay.

In one embodiment, this assay is used to determine target sites in the α-1 antitrypsin RNA target for dsRNA mediated RNAi cleavage, wherein a plurality of dsRNA constructs are screened for RNAi mediated cleavage of the α-1 antitrypsin RNA target, for example, by analyzing the assay reaction by electrophoresis of labeled target RNA, or by northern blotting, as well as by other methodology well known in the art.

In certain embodiments, a dsRNA of the invention is deemed to possess α-1 antitrypsin inhibitory activity if, e.g., a 50% reduction in α-1 antitrypsin RNA levels is observed in a system, cell, tissue or organism, relative to a suitable control. Additional metes and bounds for determination of α-1 antitrypsin inhibitory activity of a dsRNA of the invention are described supra.

Conjugation and Delivery of Anti-α-1 Antitrypsin dsRNA Agents

In certain embodiments the present invention relates to a method for treating a subject having a α-1 antitrypsin-associated disease or disorder, or at risk of developing a α-1 antitrypsin-associated disease or disorder. In such embodiments, the dsRNA can act as novel therapeutic agents for controlling the α-1 antitrypsin-associated disease or disorder. The method comprises administering a pharmaceutical composition of the invention to the patient (e.g., human), such that the expression, level and/or activity of a α-1 antitrypsin RNA is reduced. The expression, level and/or activity of a polypeptide encoded by a α-1 antitrypsin RNA might also be reduced by a dsRNA of the instant invention, even where said dsRNA is directed against a non-coding region of the α-1 antitrypsin transcript (e.g., a targeted 5′ UTR or 3′ UTR sequence). Because of their high specificity, the dsRNAs of the present invention can specifically target α-1 antitrypsin sequences of cells and tissues, optionally in an allele-specific manner where polymorphic alleles exist within an individual and/or population.

In the treatment of a α-1 antitrypsin-associated disease or disorder, the dsRNA can be brought into contact with the cells or tissue of a subject, e.g., the cells or tissue of a subject exhibiting disregulation of α-1 antitrypsin and/or otherwise targeted for reduction of α-1 antitrypsin levels. For example, dsRNA substantially identical to all or part of a α-1 antitrypsin RNA sequence, may be brought into contact with or introduced into such a cell, either in vivo or in vitro. Similarly, dsRNA substantially identical to all or part of a α-1 antitrypsin RNA sequence may administered directly to a subject having or at risk of developing a α-1 antitrypsin-associated disease or disorder.

Therapeutic use of the dsRNA agents of the instant invention can involve use of formulations of dsRNA agents comprising multiple different dsRNA agent sequences. For example, two or more, three or more, four or more, five or more, etc. of the presently described agents can be combined to produce a formulation that, e.g., targets multiple different regions of the α-1 antitrypsin RNA, or that not only target α-1 antitrypsin RNA but also target, e.g., cellular target genes associated with a α-1 antitrypsin-associated disease or disorder. A dsRNA agent of the instant invention may also be constructed such that either strand of the dsRNA agent independently targets two or more regions of α-1 antitrypsin RNA, or such that one of the strands of the dsRNA agent targets a cellular target gene of α-1 antitrypsin known in the art.

Use of multifunctional dsRNA molecules that target more then one region of a target nucleic acid molecule can also provide potent inhibition of α-1 antitrypsin RNA levels and expression. For example, a single multifunctional dsRNA construct of the invention can target both the α-1 antitrypsin-506 and α-1 antitrypsin-1059 sites simultaneously; additionally and/or alternatively, single or multifunctional agents of the invention can be designed to selectively target one splice variant of α-1 antitrypsin over another.

Thus, the dsRNA agents of the instant invention, individually, or in combination or in conjunction with other drugs, can be used to treat, inhibit, reduce, or prevent a α-1 antitrypsin-associated disease or disorder. For example, the dsRNA molecules can be administered to a subject or can be administered to other appropriate cells evident to those skilled in the art, individually or in combination with one or more drugs under conditions suitable for the treatment.

The dsRNA molecules also can be used in combination with other known treatments to treat, inhibit, reduce, or prevent a α-1 antitrypsin-associated disease or disorder in a subject or organism. For example, the described molecules could be used in combination with one or more known compounds, treatments, or procedures to treat, inhibit, reduce, or prevent a α-1 antitrypsin-associated disease or disorder in a subject or organism as are known in the art.

A dsRNA agent of the invention can be conjugated (e.g., at its 5′ or 3′ terminus of its sense or antisense strand) or unconjugated to another moiety (e.g. a non-nucleic acid moiety such as a peptide), an organic compound (e.g., a dye, cholesterol, or the like). Modifying dsRNA agents in this way may improve cellular uptake or enhance cellular targeting activities of the resulting dsRNA agent derivative as compared to the corresponding unconjugated dsRNA agent, are useful for tracing the dsRNA agent derivative in the cell, or improve the stability of the dsRNA agent derivative compared to the corresponding unconjugated dsRNA agent.

Methods of Introducing Nucleic Acids, Vectors, and Host Cells

dsRNA agents of the invention may be directly introduced into a cell (i.e., intracellularly); or introduced extracellularly into a cavity, interstitial space, into the circulation of an organism, introduced orally, or may be introduced by bathing a cell or organism in a solution containing the nucleic acid. Vascular or extravascular circulation, the blood or lymph system, and the cerebrospinal fluid are sites where the nucleic acid may be introduced.

The dsRNA agents of the invention can be introduced using nucleic acid delivery methods known in art including injection of a solution containing the nucleic acid, bombardment by particles covered by the nucleic acid, soaking the cell or organism in a solution of the nucleic acid, or electroporation of cell membranes in the presence of the nucleic acid. Other methods known in the art for introducing nucleic acids to cells may be used, such as lipid-mediated carrier transport, chemical-mediated transport, and cationic liposome transfection such as calcium phosphate, and the like. The nucleic acid may be introduced along with other components that perform one or more of the following activities: enhance nucleic acid uptake by the cell or other-wise increase inhibition of the target α-1 antitrypsin RNA.

A cell having a target α-1 antitrypsin RNA may be from the germ line or somatic, totipotent or pluripotent, dividing or non-dividing, parenchyma or epithelium, immortalized or transformed, or the like. The cell may be a stem cell or a differentiated cell. Cell types that are differentiated include adipocytes, fibroblasts, myocytes, cardiomyocytes, endothelium, neurons, glia, blood cells, megakaryocytes, lymphocytes, macrophages, neutrophils, eosinophils, basophils, mast cells, leukocytes, granulocytes, keratinocytes, chondrocytes, osteoblasts, osteoclasts, hepatocytes, and cells of the endocrine or exocrine glands.

Depending on the particular target α-1 antitrypsin RNA sequence and the dose of dsRNA agent material delivered, this process may provide partial or complete loss of function for the α-1 antitrypsin RNA. A reduction or loss of RNA levels or expression (either α-1 antitrypsin RNA expression or encoded polypeptide expression) in at least 50%, 60%, 70%, 80%, 90%, 95% or 99% or more of targeted cells is exemplary. Inhibition of α-1 antitrypsin RNA levels or expression refers to the absence (or observable decrease) in the level of α-1 antitrypsin RNA or α-1 antitrypsin RNA-encoded protein. Specificity refers to the ability to inhibit the α-1 antitrypsin RNA without manifest effects on other genes of the cell. The consequences of inhibition can be confirmed by examination of the outward properties of the cell or organism or by biochemical techniques such as RNA solution hybridization, nuclease protection, Northern hybridization, reverse transcription, gene expression monitoring with a microarray, antibody binding, enzyme linked immunosorbent assay (ELISA), Western blotting, radioimmunoassay (RIA), other immunoassays, and fluorescence activated cell analysis (FACS). Inhibition of target α-1 antitrypsin RNA sequence(s) by the dsRNA agents of the invention also can be measured based upon the effect of administration of such dsRNA agents upon development/progression of an α-1 antitrypsin-associated disease or disorder, e.g., chronic liver disease, liver inflammation, cirrhosis, liver fibrosis, and/or hepatocellular carcinoma, etc., either in vivo or in vitro. Treatment of any of these preceding liver conditions can be assessed by art-recognized tests for liver function, e.g., determination of the percentage of liver function (e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80% or greater) that is being achieved relative to average healthy liver function of an appropriate control population. In certain embodiments, successful treatment result in a decline or halting of reduction in total liver function associated with a liver disease. In some embodiments, liver function improves with successful treatment. Treatment and/or reductions in hepatocellular carcinoma tumor or cancer cell levels can include halting or reduction of growth of tumor or cancer cell levels or reductions of, e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 99% or more, and can also be measured in logarithmic terms, e.g., 10-fold, 100-fold, 1000-fold, 10⁵-fold, 10⁶-fold, 10⁷-fold reduction in cancer cell levels could be achieved via administration of the dsRNA agents of the invention to cells, a tissue, or a subject.

For RNA-mediated inhibition in a cell line or whole organism, expression a reporter or drug resistance gene whose protein product is easily assayed can be measured. Such reporter genes include acetohydroxyacid synthase (AHAS), alkaline phosphatase (AP), beta galactosidase (LacZ), beta glucoronidase (GUS), chloramphenicol acetyltransferase (CAT), green fluorescent protein (GFP), horseradish peroxidase (HRP), luciferase (Luc), nopaline synthase (NOS), octopine synthase (OCS), and derivatives thereof. Multiple selectable markers are available that confer resistance to ampicillin, bleomycin, chloramphenicol, gentamycin, hygromycin, kanamycin, lincomycin, methotrexate, phosphinothricin, puromycin, and tetracyclin. Depending on the assay, quantitation of the amount of gene expression allows one to determine a degree of inhibition which is greater than 10%, 33%, 50%, 90%, 95% or 99% as compared to a cell not treated according to the present invention.

Lower doses of injected material and longer times after administration of RNA silencing agent may result in inhibition in a smaller fraction of cells (e.g., at least 10%, 20%, 50%, 75%, 90%, or 95% of targeted cells). Quantitation of gene expression in a cell may show similar amounts of inhibition at the level of accumulation of target α-1 antitrypsin RNA or translation of target protein. As an example, the efficiency of inhibition may be determined by assessing the amount of gene product in the cell; RNA may be detected with a hybridization probe having a nucleotide sequence outside the region used for the inhibitory dsRNA, or translated polypeptide may be detected with an antibody raised against the polypeptide sequence of that region.

The dsRNA agent may be introduced in an amount which allows delivery of at least one copy per cell. Higher doses (e.g., at least 5, 10, 100, 500 or 1000 copies per cell) of material may yield more effective inhibition; lower doses may also be useful for specific applications.

Pharmaceutical Compositions

In certain embodiments, the present invention provides for a pharmaceutical composition comprising the dsRNA agent of the present invention. The dsRNA agent sample can be suitably formulated and introduced into the environment of the cell by any means that allows for a sufficient portion of the sample to enter the cell to induce gene silencing, if it is to occur. Many formulations for dsRNA are known in the art and can be used so long as the dsRNA gains entry to the target cells so that it can act. See, e.g., U.S. published patent application Nos. 2004/0203145 A1 and 2005/0054598 A1. For example, the dsRNA agent of the instant invention can be formulated in buffer solutions such as phosphate buffered saline solutions, liposomes, micellar structures, and capsids. Formulations of dsRNA agent with cationic lipids can be used to facilitate transfection of the dsRNA agent into cells. For example, cationic lipids, such as lipofectin (U.S. Pat. No. 5,705,188), cationic glycerol derivatives, and polycationic molecules, such as polylysine (published PCT International Application WO 97/30731), can be used. Suitable lipids include Oligofectamine, Lipofectamine (Life Technologies), NC388 (Ribozyme Pharmaceuticals, Inc., Boulder, Colo.), or FuGene 6 (Roche) all of which can be used according to the manufacturer's instructions.

Such compositions typically include the nucleic acid molecule and a pharmaceutically acceptable carrier. As used herein the language “pharmaceutically acceptable carrier” includes saline, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Supplementary active compounds can also be incorporated into the compositions.

A pharmaceutical composition is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL.TM. (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringability exists. It should be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyetheylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as manitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in a selected solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle, which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

Oral compositions generally include an inert diluent or an edible carrier. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules, e.g., gelatin capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, the compounds are delivered in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer. Such methods include those described in U.S. Pat. No. 6,468,798.

Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active compounds are formulated into ointments, salves, gels, or creams as generally known in the art.

The compounds can also be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

The compounds can also be administered by transfection or infection using methods known in the art, including but not limited to the methods described in McCaffrey et al. (2002), Nature, 418(6893), 38-9 (hydrodynamic transfection); Xia et al. (2002), Nature Biotechnol., 20(10), 1006-10 (viral-mediated delivery); or Putnam (1996), Am. J. Health Syst. Pharm. 53(2), 151-160, erratum at Am. J. Health Syst. Pharm. 53(3), 325 (1996).

The compounds can also be administered by a method suitable for administration of nucleic acid agents, such as a DNA vaccine. These methods include gene guns, bio injectors, and skin patches as well as needle-free methods such as the micro-particle DNA vaccine technology disclosed in U.S. Pat. No. 6,194,389, and the mammalian transdermal needle-free vaccination with powder-form vaccine as disclosed in U.S. Pat. No. 6,168,587. Additionally, intranasal delivery is possible, as described in, inter alia, Hamajima et al. (1998), Clin. Immunol. Immunopathol., 88(2), 205-10. Liposomes (e.g., as described in U.S. Pat. No. 6,472,375) and microencapsulation can also be used. Biodegradable targetable microparticle delivery systems can also be used (e.g., as described in U.S. Pat. No. 6,471,996).

In one embodiment, the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Such formulations can be prepared using standard techniques. The materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811.

Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD₅₀ (the dose lethal to 50% of the population) and the ED₅₀ (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD₅₀/ED₅₀. Compounds which exhibit high therapeutic indices are preferred. While compounds that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissue in order to minimize potential damage to uninfected cells and, thereby, reduce side effects.

The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED₅₀ with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For a compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the IC₅₀ (i.e., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography.

As defined herein, a therapeutically effective amount of a nucleic acid molecule (i.e., an effective dosage) depends on the nucleic acid selected. For instance, single dose amounts of a dsRNA (or, e.g., a construct(s) encoding for such dsRNA) in the range of approximately 1 pg to 1000 mg may be administered; in some embodiments, 10, 30, 100, or 1000 pg, or 10, 30, 100, or 1000 ng, or 10, 30, 100, or 1000 μg, or 10, 30, 100, or 1000 mg may be administered. In some embodiments, 1-5 g of the compositions can be administered. The compositions can be administered one from one or more times per day to one or more times per week; including once every other day. The skilled artisan will appreciate that certain factors may influence the dosage and timing required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and other diseases present. Moreover, treatment of a subject with a therapeutically effective amount of a nucleic acid (e.g., dsRNA), protein, polypeptide, or antibody can include a single treatment or, preferably, can include a series of treatments.

The nucleic acid molecules of the invention can be inserted into expression constructs, e.g., viral vectors, retroviral vectors, expression cassettes, or plasmid viral vectors, e.g., using methods known in the art, including but not limited to those described in Xia et al., (2002), supra. Expression constructs can be delivered to a subject by, for example, inhalation, orally, intravenous injection, local administration (see U.S. Pat. No. 5,328,470) or by stereotactic injection (see e.g., Chen et al. (1994), Proc. Natl. Acad. Sci. USA, 91, 3054-3057). The pharmaceutical preparation of the delivery vector can include the vector in an acceptable diluent, or can comprise a slow release matrix in which the delivery vehicle is imbedded. Alternatively, where the complete delivery vector can be produced intact from recombinant cells, e.g., retroviral vectors, the pharmaceutical preparation can include one or more cells which produce the gene delivery system.

The expression constructs may be constructs suitable for use in the appropriate expression system and include, but are not limited to retroviral vectors, linear expression cassettes, plasmids and viral or virally-derived vectors, as known in the art. Such expression constructs may include one or more inducible promoters, RNA Pol III promoter systems such as U6 snRNA promoters or H1 RNA polymerase III promoters, or other promoters known in the art. The constructs can include one or both strands of the siRNA. Expression constructs expressing both strands can also include loop structures linking both strands, or each strand can be separately transcribed from separate promoters within the same construct. Each strand can also be transcribed from a separate expression construct, e.g., Tuschl (2002, Nature Biotechnol 20: 500-505).

It can be appreciated that the method of introducing dsRNA agents into the environment of the cell will depend on the type of cell and the make up of its environment. For example, when the cells are found within a liquid, one preferable formulation is with a lipid formulation such as in lipofectamine and the dsRNA agents can be added directly to the liquid environment of the cells. Lipid formulations can also be administered to animals such as by intravenous, intramuscular, or intraperitoneal injection, or orally or by inhalation or other methods as are known in the art. When the formulation is suitable for administration into animals such as mammals and more specifically humans, the formulation is also pharmaceutically acceptable. Pharmaceutically acceptable formulations for administering oligonucleotides are known and can be used. In some instances, it may be preferable to formulate dsRNA agents in a buffer or saline solution and directly inject the formulated dsRNA agents into cells, as in studies with oocytes. The direct injection of dsRNA agent duplexes may also be done. For suitable methods of introducing dsRNA (e.g., DsiRNA agents), see U.S. published patent application No. 2004/0203145 A1.

Suitable amounts of a dsRNA agent must be introduced and these amounts can be empirically determined using standard methods. Typically, effective concentrations of individual dsRNA agent species in the environment of a cell will be 50 nanomolar or less, 10 nanomolar or less, or compositions in which concentrations of 1 nanomolar or less can be used. In another embodiment, methods utilizing a concentration of 200 picomolar or less, 100 picomolar or less, 50 picomolar or less, 20 picomolar or less, and even a concentration of 10 picomolar or less, 5 picomolar or less, 2 picomolar or less or 1 picomolar or less can be used in many circumstances.

The method can be carried out by addition of the dsRNA agent compositions to an extracellular matrix in which cells can live provided that the dsRNA agent composition is formulated so that a sufficient amount of the dsRNA agent can enter the cell to exert its effect. For example, the method is amenable for use with cells present in a liquid such as a liquid culture or cell growth media, in tissue explants, or in whole organisms, including animals, such as mammals and especially humans.

The level or activity of a α-1 antitrypsin RNA can be determined by a suitable method now known in the art or that is later developed. It can be appreciated that the method used to measure a target RNA and/or the expression of a target RNA can depend upon the nature of the target RNA. For example, where the target α-1 antitrypsin RNA sequence encodes a protein, the term “expression” can refer to a protein or the α-1 antitrypsin RNA/transcript derived from the α-1 antitrypsin gene (either genomic or of exogenous origin). In such instances the expression of the target α-1 antitrypsin RNA can be determined by measuring the amount of α-1 antitrypsin RNA/transcript directly or by measuring the amount of α-1 antitrypsin protein. Protein can be measured in protein assays such as by staining or immunoblotting or, if the protein catalyzes a reaction that can be measured, by measuring reaction rates. All such methods are known in the art and can be used. Where target α-1 antitrypsin RNA levels are to be measured, art-recognized methods for detecting RNA levels can be used (e.g., RT-PCR, Northern Blotting, etc.). In targeting α-1 antitrypsin RNAs with the dsRNA agents of the instant invention, it is also anticipated that measurement of the efficacy of a dsRNA agent in reducing levels of α-1 antitrypsin RNA or protein in a subject, tissue, in cells, either in vitro or in vivo, or in cell extracts can also be used to determine the extent of reduction of α-1 antitrypsin-associated phenotypes (e.g., disease or disorders, e.g., chronic liver disease, liver inflammation, cirrhosis, liver fibrosis, and/or hepatocellular carcinoma, etc.). The above measurements can be made on cells, cell extracts, tissues, tissue extracts or other suitable source material.

The determination of whether the expression of a α-1 antitrypsin RNA has been reduced can be by a suitable method that can reliably detect changes in RNA levels. Typically, the determination is made by introducing into the environment of a cell undigested dsRNA such that at least a portion of that dsRNA agent enters the cytoplasm, and then measuring the level of the target RNA. The same measurement is made on identical untreated cells and the results obtained from each measurement are compared.

The dsRNA agent can be formulated as a pharmaceutical composition which comprises a pharmacologically effective amount of a dsRNA agent and pharmaceutically acceptable carrier. A pharmacologically or therapeutically effective amount refers to that amount of a dsRNA agent effective to produce the intended pharmacological, therapeutic or preventive result. The phrases “pharmacologically effective amount” and “therapeutically effective amount” or simply “effective amount” refer to that amount of an RNA effective to produce the intended pharmacological, therapeutic or preventive result. For example, if a given clinical treatment is considered effective when there is at least a 20% reduction in a measurable parameter associated with a disease or disorder, a therapeutically effective amount of a drug for the treatment of that disease or disorder is the amount necessary to effect at least a 20% reduction in that parameter.

Suitably formulated pharmaceutical compositions of this invention can be administered by means known in the art such as by parenteral routes, including intravenous, intramuscular, intraperitoneal, subcutaneous, transdermal, airway (aerosol), rectal, vaginal and topical (including buccal and sublingual) administration. In some embodiments, the pharmaceutical compositions are administered by intravenous or intraparenteral infusion or injection.

In general, a suitable dosage unit of dsRNA will be in the range of 0.001 to 0.25 milligrams per kilogram body weight of the recipient per day, or in the range of 0.01 to 20 micrograms per kilogram body weight per day, or in the range of 0.001 to 5 micrograms per kilogram of body weight per day, or in the range of 1 to 500 nanograms per kilogram of body weight per day, or in the range of 0.01 to 10 micrograms per kilogram body weight per day, or in the range of 0.10 to 5 micrograms per kilogram body weight per day, or in the range of 0.1 to 2.5 micrograms per kilogram body weight per day. A pharmaceutical composition comprising the dsRNA can be administered once daily. However, the therapeutic agent may also be dosed in dosage units containing two, three, four, five, six or more sub-doses administered at appropriate intervals throughout the day. In that case, the dsRNA contained in each sub-dose must be correspondingly smaller in order to achieve the total daily dosage unit. The dosage unit can also be compounded for a single dose over several days, e.g., using a conventional sustained release formulation which provides sustained and consistent release of the dsRNA over a several day period. Sustained release formulations are well known in the art. In this embodiment, the dosage unit contains a corresponding multiple of the daily dose. Regardless of the formulation, the pharmaceutical composition must contain dsRNA in a quantity sufficient to inhibit expression of the target gene in the animal or human being treated. The composition can be compounded in such a way that the sum of the multiple units of dsRNA together contain a sufficient dose.

Data can be obtained from cell culture assays and animal studies to formulate a suitable dosage range for humans. The dosage of compositions of the invention lies within a range of circulating concentrations that include the ED₅₀ (as determined by known methods) with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For a compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range of the compound that includes the IC₅₀ (i.e., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels of dsRNA in plasma may be measured by standard methods, for example, by high performance liquid chromatography.

The pharmaceutical compositions can be included in a kit, container, pack, or dispenser together with instructions for administration.

Methods of Treatment

The present invention provides for both prophylactic and therapeutic methods of treating a subject at risk of (or susceptible to) a disease or disorder caused, in whole or in part, by α-1 antitrypsin (e.g., misregulation and/or elevation of α-1 antitrypsin transcript and/or α-1 antitrypsin protein levels), or treatable via selective targeting of α-1 antitrypsin.

In certain aspects, the invention provides a method for preventing in a subject, a disease or disorder as described herein (including, e.g., prevention of the commencement of transforming events within a subject via inhibition of α-1 antitrypsin expression), by administering to the subject a therapeutic agent (e.g., a dsRNA agent or vector or transgene encoding same). Subjects at risk for the disease can be identified by, for example, one or a combination of diagnostic or prognostic assays as described herein. Administration of a prophylactic agent can occur prior to the detection of, e.g., cancer in a subject, or the manifestation of symptoms characteristic of the disease or disorder, such that the disease or disorder is prevented or, alternatively, delayed in its progression.

Another aspect of the invention pertains to methods of treating subjects therapeutically, i.e., altering the onset of symptoms of the disease or disorder. These methods can be performed in vitro (e.g., by culturing the cell with the dsRNA agent) or, alternatively, in vivo (e.g., by administering the dsRNA agent to a subject).

With regard to both prophylactic and therapeutic methods of treatment, such treatments may be specifically tailored or modified, based on knowledge obtained from the field of pharmacogenomics. “Pharmacogenomics”, as used herein, refers to the application of genomics technologies such as gene sequencing, statistical genetics, and gene expression analysis to drugs in clinical development and on the market. More specifically, the term refers the study of how a patient's genes determine his or her response to a drug (e.g., a patient's “drug response phenotype”, or “drug response genotype”). Thus, another aspect of the invention provides methods for tailoring an individual's prophylactic or therapeutic treatment with either the target α-1 antitrypsin RNA molecules of the present invention or target α-1 antitrypsin RNA modulators according to that individual's drug response genotype. Pharmacogenomics allows a clinician or physician to target prophylactic or therapeutic treatments to patients who will most benefit from the treatment and to avoid treatment of patients who will experience toxic drug-related side effects.

The practice of the present invention employs, unless otherwise indicated, conventional techniques of chemistry, molecular biology, microbiology, recombinant DNA, genetics, immunology, cell biology, cell culture and transgenic biology, which are within the skill of the art. See, e.g., Maniatis et al., 1982, Molecular Cloning (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.); Sambrook et al., 1989, Molecular Cloning, 2nd Ed. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.); Sambrook and Russell, 2001, Molecular Cloning, 3rd Ed. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.); Ausubel et al., 1992), Current Protocols in Molecular Biology (John Wiley & Sons, including periodic updates); Glover, 1985, DNA Cloning (IRL Press, Oxford); Anand, 1992; Guthrie and Fink, 1991; Harlow and Lane, 1988, Antibodies, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.); Jakoby and Pastan, 1979; Nucleic Acid Hybridization (B. D. Hames & S. J. Higgins eds. 1984); Transcription And Translation (B. D. Hames & S. J. Higgins eds. 1984); Culture Of Animal Cells (R. I. Freshney, Alan R. Liss, Inc., 1987); Immobilized Cells And Enzymes (IRL Press, 1986); B. Perbal, A Practical Guide To Molecular Cloning (1984); the treatise, Methods In Enzymology (Academic Press, Inc., N.Y.); Gene Transfer Vectors For Mammalian Cells (J. H. Miller and M. P. Calos eds., 1987, Cold Spring Harbor Laboratory); Methods In Enzymology, Vols. 154 and 155 (Wu et al. eds.), Immunochemical Methods In Cell And Molecular Biology (Mayer and Walker, eds., Academic Press, London, 1987); Handbook Of Experimental Immunology, Volumes I-IV (D. M. Weir and C. C. Blackwell, eds., 1986); Riott, Essential Immunology, 6th Edition, Blackwell Scientific Publications, Oxford, 1988; Hogan et al., Manipulating the Mouse Embryo, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1986); Westerfield, M., The zebrafish book. A guide for the laboratory use of zebrafish (Danio rerio), (4th Ed., Univ. of Oregon Press, Eugene, 2000).

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

EXAMPLES

The present invention is described by reference to the following Examples, which are offered by way of illustration and are not intended to limit the invention in any manner. Standard techniques well known in the art or the techniques specifically described below were utilized.

Example 1: Preparation of Double-Stranded RNA Oligonucleotides

Oligonucleotide Synthesis and Purification

DsiRNA molecules can be designed to interact with various sites in the RNA message, for example, target sequences within the RNA sequences described herein. In presently exemplified agents, 384 human target α-1 antitrypsin sequences were selected for evaluation (a selection of the 384 human target α-1 antitrypsin sites were predicted to be conserved with corresponding sites in the mouse α-1 antitrypsin transcript sequence). The sequences of one strand of the DsiRNA molecules were complementary to the target α-1 antitrypsin site sequences described above. The DsiRNA molecules were chemically synthesized using methods described herein. Generally, DsiRNA constructs were synthesized using solid phase oligonucleotide synthesis methods as described for 19-23mer siRNAs (see for example Usman et al., U.S. Pat. Nos. 5,804,683; 5,831,071; 5,998,203; 6,117,657; 6,353,098; 6,362,323; 6,437,117; 6,469,158; Scaringe et al., U.S. Pat. Nos. 6,111,086; 6,008,400; 6,111,086).

Individual RNA strands were synthesized and HPLC purified according to standard methods (Integrated DNA Technologies, Coralville, Iowa). For example, RNA oligonucleotides were synthesized using solid phase phosphoramidite chemistry, deprotected and desalted on NAP-5 columns (Amersham Pharmacia Biotech, Piscataway, N.J.) using standard techniques (Damha and Olgivie, 1993, Methods Mol Biol 20: 81-114; Wincott et al., 1995, Nucleic Acids Res 23: 2677-84). The oligomers were purified using ion-exchange high performance liquid chromatography (IE-HPLC) on an Amersham Source 15Q column (1.0 cm×25 cm; Amersham Pharmacia Biotech, Piscataway, N.J.) using a 15 min step-linear gradient. The gradient varied from 90:10 Buffers A:B to 52:48 Buffers A:B, where Buffer A was 100 mM Tris pH 8.5 and Buffer B was 100 mM Tris pH 8.5, 1 M NaCl. Samples were monitored at 260 nm and peaks corresponding to the full-length oligonucleotide species were collected, pooled, desalted on NAP-5 columns, and lyophilized.

The purity of each oligomer was determined by capillary electrophoresis (CE) on a Beckman PACE 5000 (Beckman Coulter, Inc., Fullerton, Calif.). The CE capillaries had a 100 μm inner diameter and contained ssDNA 100R Gel (Beckman-Coulter). Typically, about 0.6 nmole of oligonucleotide was injected into a capillary, run in an electric field of 444 V/cm and detected by UV absorbance at 260 nm. Denaturing Tris-Borate-7 M-urea running buffer was purchased from Beckman-Coulter. Oligoribonucleotides were obtained that were at least 90% pure as assessed by CE for use in experiments described below. Compound identity was verified by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectroscopy on a Voyager DE.TM. Biospectometry Work Station (Applied Biosystems, Foster City, Calif.) following the manufacturer's recommended protocol. Relative molecular masses of all oligomers were obtained, often within 0.2% of expected molecular mass.

Preparation of Duplexes

Single-stranded RNA (ssRNA) oligomers were resuspended, e.g., at 100 μM concentration in duplex buffer consisting of 100 mM potassium acetate, 30 mM HEPES, pH 7.5. Complementary sense and antisense strands were mixed in equal molar amounts to yield a final solution of, e.g., 50 μM duplex. Samples were heated to 100° C. for 5′ in RNA buffer (IDT) and allowed to cool to room temperature before use. Double-stranded RNA (dsRNA) oligomers were stored at −20° C. Single-stranded RNA oligomers were stored lyophilized or in nuclease-free water at −80° C.

Nomenclature

For consistency, the following nomenclature has been employed in the instant specification. Names given to duplexes indicate the length of the oligomers and the presence or absence of overhangs. A “25/27” is an asymmetric duplex having a 25 base sense strand and a 27 base antisense strand with a 2-base 3′-overhang. A “27/25” is an asymmetric duplex having a 27 base sense strand and a 25 base antisense strand.

Cell Culture and RNA Transfection

Huh7 cells were obtained and maintained in DMEM (HyClone) supplemented with 10% fetal bovine serum (HyClone) at 37° C. under 5% CO₂. For RNA transfections, cells were transfected with DsiRNAs at a final concentration of 1 nM, 0.1 nM or 0.03 nM using Lipofectamine™ RNAiMAX (Invitrogen) and following manufacturer's instructions. Briefly, for 0.1 nM transfections, e.g., of Example 3 below, an aliquot of stock solution of each DsiRNA was mixed with Opti-MEM I (Invitrogen) and Lipofectamine™ RNAiMAX to reach a volume of 150 μL (with 0.3 nM DsiRNA). The resulting 150 μL mix was incubated for 20 min at RT to allow DsiRNA:Lipofectamine™ RNAiMAX complexes to form. Meanwhile, target cells were trypsinized and resuspended in medium. At the end of the 20 min of complexation, 50 uL of the DsiRNA:RNAiMAX mixture was added per well into triplicate wells of 96 well plates. Finally, 100 μL of the cell suspension was added to each well (final volume 150 μL) and plates were placed into the incubator for 24 hours.

Assessment of α-1 Antitrypsin Inhibition

-   -   α-1 antitrypsin target gene knockdown was determined by qRT-PCR,         with values normalized to HPRT and SFRS9 housekeeping genes, and         to transfections with control DsiRNAs and/or mock transfection         controls.

RNA Isolation and Analysis

Media was aspirated, and total RNA was extracted using the SV96 kit (Promega). Total RNA was reverse-transcribed using SuperscriptII, Oligo dT, and random hexamers following manufacturer's instructions. Typically, the resulting cDNA was analyzed by qPCR using primers and probes specific for both the α-1 antitrypsin gene and for the human genes HPRT-1 and SFRS9. An ABI 7700 was used for the amplification reactions. Each sample was tested in triplicate. Relative α-1 antitrypsin RNA levels were normalized to HPRT1 and SFRS9 RNA levels and compared with RNA levels obtained in transfection control samples.

Example 2: DsiRNA Inhibition of α-1 Antitrypsin

DsiRNA molecules targeting α-1 antitrypsin were designed and synthesized as described above and tested in human Huh7 cells (alternatively, HepG2 or other human cells could have been used) for inhibitory efficacy. For transfection, annealed DsiRNAs were mixed with the transfection reagent (Lipofectamine™ RNAiMAX, Invitrogen) and incubated for 20 minutes at room temperature. The Huh7 (human) or AML12 (mouse) cells (alternatively, mouse Hepa 1-6 or other mouse cells could have been used) were trypsinized, resuspended in media, and added to wells (100 uL per well) to give a final DsiRNA concentration of 1 nM in a volume of 150 μl. Each DsiRNA transfection mixture was added to 3 wells for triplicate DsiRNA treatments. Cells were incubated at 37° C. for 24 hours in the continued presence of the DsiRNA transfection mixture. At 24 hours, RNA was prepared from each well of treated cells. The supernatants with the transfection mixtures were first removed and discarded, then the cells were lysed and RNA was prepared from each well. Target α-1 antitrypsin RNA levels following treatment were evaluated by qRT-PCR for the α-1 antitrypsin target gene, with values normalized to those obtained for controls. Triplicate data was averaged and the % error was determined for each treatment. Normalized data were both tablulated and graphed, and the reduction of target mRNA by active DsiRNAs in comparison to controls was determined (see Table 22 below and FIGS. 2A to 2D).

TABLE 22 α-1 Antitrypsin Inhibitory Efficacy of DsiRNAs Assayed at 1 nM in Human Huh7 and Mouse AML12 Cells Human-Huh7 Mouse-AML12 Normalized HPRT/SFRS9; Normalized HPRT/Rpl23; vs NC1, NC5, NC7 vs NC1, NC5, NC7 Hs 462-563 Hs 1811-1910 Mm 331-462 Mm 1532-1462 Duplex Mm Rhesus (FAM) Assay (HEX) Assay (FAM) Assay (HEX) Assay Name Location Location % Remaining % Remaining % Remaining % Remaining AAT-364 335 AAT-366 337 AAT-367 338 11.5 ± 8.7   58.7 ± 22.9 AAT-368 339 6.9 ± 14.4 67.2 ± 28.6 AAT-369 340 5.3 ± 21.7 81.5 ± 25.7 AAT-370 341 38.2 ± 15    70.3 ± 21.6 AAT-371 342 8.7 ± 3.2    82 ± 28.2 AAT-391 362 9.8 ± N/A 60.8 ± N/A AAT-392 363 AAT-393 364 AAT-394 365 12.3 ± 13.1  33.8 ± 29.8 AAT-395 366 2.6 ± 3.3  23.6 ± 7.1  AAT-475 446 2.5 ± 10.2 18.8 ± 3.6  AAT-477 448 6.7 ± 11.5 38.1 ± 13.1 AAT-480 451 7.2 ± 8.1  38.9 ± 14.2 AAT-481 452 12.1 ± 6.1   40.7 ± 13.1 AAT-482 453 AAT-483 454 10.2 ± 18.9  26.5 ± 21.2 AAT-484 455 7.2 ± 8.8  22.1 ± 7.7  AAT-485 456 6.6 ± 9.5  20.6 ± 19.8 AAT-486 457 1.6 ± 3.3  18.5 ± 5.5  AAT-487 458 7.8 ± 2.3  33.1 ± 7.9  AAT-488 459 24.5 ± 7.8     45 ± 12.8 AAT-489 460 27.4 ± 8.9   54.3 ± 6.1  AAT-490 461 2.6 ± 27.3 30.7 ± 9.3  AAT-491 462 2.6 ± 15.9 25 ± 8.2 AAT-492 463 3.5 ± 5    25.4 ± 6.5  AAT-493 464 6.6 ± 20.1 34.6 ± 3.8  AAT-494 465 18.5 ± 5     42.2 ± 8.2  AAT-495 466 7.2 ± 5    31.3 ± 9.2  AAT-496 467 1.3 ± 1.7  21.6 ± 5.1  AAT-497 468 7.3 ± 11.9 30.3 ± 10.9 AAT-498 469 42.4 ± 4.1   64.8 ± 6.7  AAT-499 470 40.2 ± 3     59 ± 3.6 AAT-500 471 7.9 ± 5.6  26.3 ± 3.1  AAT-501 472 9.4 ± 3.5  31.8 ± 8.6  AAT-502 473 11.5 ± 10.4  28.5 ± 7.9  AAT-503 474 21.5 ± 6.4   38.3 ± 11   AAT-504 475 13.3 ± 7.8   36.1 ± 8.4  AAT-505 328 17.9 ± 6.6   42.3 ± 3.1  40.4 ± 2.7 43.3 ± 5.2 AAT-506 329 33.5 ± 3.3   54.9 ± 7.5  51.7 ± 7.9 55.8 ± 8.6 AAT-507 330 26.5 ± 3.5    48 ± 3.8 56.8 ± 2.1 59.2 ± 2.9 AAT-508 331 10 ± 5.5  35.3 ± 5.3    26 ± 7.5 30.2 ± 5.5 AAT-509 332 75.6 ± 2.1   98.3 ± 6.2  75.3 ± 5.1 77.3 ± 4   AAT-510 481 88.4 ± 2.1   105.1 ± 3.8   AAT-512 483 31.6 ± 1.2   62.6 ± 4    AAT-513 484 13.1 ± 5.5   27.4 ± 2.4  AAT-515 486 27 ± 6.8  44.9 ± 5.6  AAT-516 487 43 ± 7   63.6 ± 17.4 AAT-517 488 22.8 ± 4.9   38.6 ± 4.4  AAT-518 489 20.7 ± 1.4   37.2 ± 5.8  AAT-519 490 9.5 ± 7.6  24 ± 2  AAT-520 491 43.9 ± 1.7   63.8 ± 10.1 AAT-521 492 7.4 ± 12.8 52.9 ± 15.8 AAT-522 493 35.6 ± 4.9   77.6 ± 15.8 AAT-523 494 48.6 ± 9.4   98.5 ± 17.4 AAT-524 495 54.9 ± 8.9   122.3 ± 12.2  AAT-525 496 20.3 ± 28.7  53.4 ± 23.3 AAT-526 497 5.2 ± 27.3 58.2 ± 7.1  AAT-527 498 24.4 ± 13.3  81.1 ± 9.9  AAT-528 499 26.2 ± 13    82.3 ± 6.1  AAT-529 500 53.4 ± 12.9  91.7 ± 26.6 AAT-530 501 40.8 ± 14.3  77.3 ± 23.4 AAT-531 502 22.3 ± 10.5  53 ± 7  AAT-532 503 11.4 ± 6     39.5 ± 18   AAT-535 506 50 ± 9.8  59.5 ± 18.1 AAT-540 511 10.6 ± 6.6     34 ± 15.5 AAT-541 512 57.2 ± 1.8    82 ± 0.4 AAT-552 523 10.4 ± 15.3  51.4 ± 7.7  AAT-555 526 71.2 ± 8.8   96.6 ± 15.2 AAT-556 527 3.6 ± 12.8   35 ± 11.1 AAT-557 528   4 ± 11.6 28.5 ± 8.7  AAT-558 529 12.8 ± 8.2   26.9 ± 5.3  AAT-579 550 7.9 ± 6.4  32.8 ± 11   AAT-580 551 28.1 ± 3.4   46.1 ± 5.4  AAT-581 552   3 ± 14.9   29 ± 13.4 AAT-582 553 12.5 ± 16    47.8 ± 6.9  AAT-583 554 5.9 ± 14.5 22.2 ± 26.5 AAT-584 555 12.9 ± 15    28.6 ± 21.8 AAT-585 556 31.9 ± 18.8  54.7 ± 23.3 AAT-586 557 3.3 ± 26.3 28.8 ± 19.8 AAT-587 558 5.6 ± 24   24.9 ± 31.1 AAT-632 603 13.7 ± 30.4  35.9 ± 28.4 AAT-633 604 43.2 ± 7.1   66.4 ± 15.6 AAT-634 605 17.2 ± 12.5  42.4 ± 7.5  AAT-637 608 84.6 ± 2.8   89.5 ± 7.6  AAT-638 609 16.8 ± 10.4  24.1 ± 14.3 AAT-671 642 39.2 ± 8.7     53 ± 12.8 AAT-673 644  8 ± 5.1 22.2 ± 6.4  AAT-674 645 27.9 ± 1.4   39.5 ± 2.4  AAT-675 646 11.2 ± 5.9   24.4 ± 14.5 AAT-676 647 92.1 ± 3.2   116.5 ± 5.4   AAT-734 705 74.3 ± 4.5   94.5 ± 3.5  AAT-735 706 8.4 ± 11.2 17.1 ± 21.3 AAT-736 707 5.1 ± 8.8   9.6 ± 12.1 AAT-737 708 2.8 ± 12.5 12.9 ± 4.4  AAT-738 709 8.3 ± 4.9  21 ± 14 AAT-739 710 75.3 ± 9.1   79.8 ± 15.3 AAT-740 711  4 ± 4.5 12.6 ± 7.2  AAT-767 738 3.5 ± 9.2  18.9 ± 12.4 AAT-768 739 57.5 ± 8.4   79.3 ± 8.9  AAT-801 772 56.1 ± 5.3   64 ± 5.3 AAT-802 773 12.8 ± 8.2   31.5 ± 5.5  AAT-803 774 8.4 ± 22.1 29.2 ± 34.4 AAT-804 775 42.7 ± 17.7  72.7 ± 17.6 AAT-805 776 6.7 ± 28   33.3 ± 26.4 AAT-806 777 13.5 ± 20.9  35.2 ± 11.3 AAT-807 778 5.8 ± 11.5 32.3 ± 5.8  AAT-808 779 33.6 ± 6.5   71.8 ± 11.4 AAT-809 780 30.7 ± 3.3   44.8 ± 5.8  AAT-810 781 2.5 ± 11.5 16.7 ± 7.5  AAT-811 782 10.4 ± 6.9   21.5 ± 4.2  AAT-812 783 2.4 ± 8.3  11.5 ± 6    AAT-813 784 15.3 ± 5.3   31.1 ± 3.4  AAT-850 821 28.1 ± 9.1   42.4 ± 7    AAT-851 822 26.4 ± 3.7   44.6 ± 7.2  AAT-852 823 22.7 ± 4.3   52.9 ± 9.8  AAT-853 824 4.2 ± 2.9  13.3 ± 19.9 AAT-854 825 27.8 ± 6.7   39.1 ± 14.2 AAT-855 826 6.9 ± 7.4  20.3 ± 17.6 AAT-856 827 11.3 ± 4.9   20.4 ± 7.4  AAT-857 828 5.1 ± 4.1  17.9 ± 5.4  AAT-858 829 8 ± 9.8 16.2 ± 20.5 AAT-859 830 5 ± 1.9  23 ± 9.3 AAT-860 831 58.4 ± 4.4   78.7 ± 10.1 AAT-861 832 83.5 ± 7.1   98.7 ± 4.3  AAT-862 833 5.5 ± 30.4 22.3 ± 29.8 AAT-863 834 21.1 ± 15.3  39.2 ± 14.1 AAT-864 835 72.9 ± 4.5   107.3 ± 7     AAT-865 836 29.1 ± 17.1  52 ± 16 AAT-866 837 7.3 ± 15.3 25.3 ± 4.7  AAT-867 838 32.1 ± 11.2  52.8 ± 9    AAT-868 839 43.1 ± 29.3  78.2 ± 17.5 AAT-869 840 10 ± 2.3  16.1 ± 5.5  AAT-870 841 23.9 ± 8.7   35.4 ± 8.5  AAT-871 842 73.3 ± 3.3   85.9 ± 2.6  AAT-872 843 13.4 ± 2.8   29.7 ± 5.9  AAT-896 867   5 ± 14.6 20.3 ± 6.8  AAT-897 868 4.2 ± 18   18.3 ± 10.1 AAT-898 869 1.5 ± 5.2  13.4 ± 5.1  AAT-899 870 3.8 ± 28.3 21.7 ± 15.8 AAT-900 871 4.6 ± 4.9  11.6 ± 8.3  AAT-901 872 4.4 ± 4    14.8 ± 17.4 AAT-902 873 5.2 ± 1.4  16.5 ± 0.9  AAT-903 874 18.3 ± 5.9   35.8 ± 9.1  AAT-904 875 20.7 ± 3.8   37.2 ± 2.4  AAT-905 876 4.8 ± 3.3  20.5 ± 5    AAT-906 877 3.3 ± 8.1  18.4 ± 2.9  AAT-907 878  11 ± 19.1 36.3 ± 17.4 AAT-908 879 21.3 ± 19.7  41.2 ± 16.7 AAT-909 880 7.5 ± 17.9 32.8 ± 9.1  AAT-910 881 5.7 ± 24.9 28.8 ± 11.8 AAT-911 882 4.4 ± 30   28.3 ± 27.3 AAT-912 883 15.1 ± 13.2  36.7 ± 15.3 AAT-913 884 11.6 ± 18.8  33.9 ± 11.3 AAT-914 885 26 ± 24  53.8 ± 14.1 AAT-915 886 30.2 + 6.8   62.1 ± 10.3 AAT-916 887 17.6 ± 11    35.3 ± 10.5 AAT-917 888 10.4 ± 8.7   28.7 ± 12.6 AAT-918 889 36.6 ± 5.3   53.2 ± 2.8  AAT-919 890 64.9 ± 1.7   88.7 ± 3.2  AAT-922 893 11.4 ± 5.9   26.5 ± 4.9  AAT-924 895 42 ± 4.9  58.2 ± 1.7  AAT-926 897 102.4 ± 3.3    132 ± 6   AAT-927 898 113 ± 1.6   143.8 ± 6.1   AAT-928 899 15.2 ± 37    27.8 ± 23.2 AAT-929 900 21.5 ± 1.9    38 ± 3.7 AAT-930 901 49.6 ± 5     60.6 ± 2.3  AAT-931 902 50.3 ± 6.9   82.5 ± 12.2 AAT-932 903 77.5 ± 10.1   98 ± 4.3 AAT-933 904 21.8 ± 7.5   41.2 ± 8.7  AAT-934 905 38.3 ± 3.7   55.3 ± 7.5  AAT-935 906  13 ± 12.7 32.6 ± 7.1  AAT-968 939  16 ± 14.4 33.2 ± 17.8 AAT-969 940 104.5 ± 5.3    141.3 ± 4.4   AAT-970 941 10.9 ± 19.6  26.6 ± 13.8 AAT-971 942 10.2 ± 11.5  30.8 ± 6.2  AAT-973 944 52.9 ± 12.8    78 ± 15.9 AAT-974 945 57.3 ± 8.1   80.5 ± 7.5  AAT-976 947 71.4 ± 9.7   101 ± 9.8  AAT-1023 994 17.2 ± 8.5   44.3 ± 10.3 AAT-1024 995 11.5 ± 6.9   26.6 ± 13   AAT-1025 996 10.2 ± 7.2   21.2 ± 9.5  AAT-1026 997 76.8 ± 2.7   98.8 ± 3.2  AAT-1055 1026 7.6 ± 2.7  22.9 ± 6.8  AAT-1059 882 93.5 ± 3.4   119.7 ± 8.9   110.7 ± 5    103.7 ± 3.5  AAT-1060 883 25.3 ± 5.5   62.7 ± 5.2  70.8 ± 3.2 61.5 ± 6.9 AAT-1061 884 61.8 ± 2.1   110.2 ± 5.2   98.4 ± 4.2 91.4 ± 2.2 AAT-1062 885 72.2 ± 4.6   90.1 ± 7.3    97 ± 4.3 95.9 ± 3   AAT-1063 886 24.7 ± 5.4   43.5 ± 7.5    62 ± 5.2 63.7 ± 3.1 AAT-1064 887 55.5 ± 7     72.8 ± 4.2  87.6 ± 6.7 90.9 ± 7.1 AAT-1065 888 23.2 ± 5.2   48.1 ± 3    60.8 ± 2.1 67.4 ± 2.9 AAT-1066 1037 9.1 ± 3.2  24.6 ± 3    AAT-1067 1038 12.8 ± 3.9    32 ± 7.5 AAT-1068 1039 47 ± 7.5  62.7 ± 6.9  AAT-1069 1040 8.2 ± 9.1  28.5 ± 10.8 AAT-1070 1041 11 ± 4.6  20.8 ± 19.1 AAT-1072 1043 65.1 ± 2.4   77.9 ± 5.5  AAT-1073 1044 15 ± 7.9  26.7 ± 3.8  AAT-1074 1045 47.7 ± 1.1   71.1 ± 1.1  AAT-1075 1046 20.2 ± N/A  29.7 ± N/A AAT-1076 1047 82.6 ± 1.9   94.9 ± 2.1  AAT-1077 1048 11.9 ± 2.6    26 ± 7.8 AAT-1078 1049 16.6 ± 3.6   42.5 ± 1.3  AAT-1079 1050 40.3 ± 13.8  74.4 ± 8.2  AAT-1080 1051 21.9 ± 7.7     60 ± 12.9 AAT-1081 1052 43.5 ± 15.7  64.8 ± 13.4 AAT-1083 1054 87.1 ± 9.3   124.7 ± 12.6  AAT-1095 1066 15.3 ± 16.2  44.6 ± 23.1 AAT-1096 1067 5.1 ± 17.4 34.2 ± 27.5 AAT-1097 1068 53.1 ± 9.1   76.4 ± 17.7 AAT-1099 1070 99.1 ± 4.1   119.5 ± 11.4  AAT-1100 1071 74.9 ± 4.7   95.6 ± 6.9  AAT-1101 1072 39.8 ± 6.8   55.1 ± 13.9 AAT-1102 1073 18.5 ± 1.6   30.8 ± 4.8  AAT-1103 1074 35.9 ± 4.2   47.3 ± 3.7  AAT-1104 1075 8.3 ± 5.7  25.9 ± 11.5 AAT-1105 1076 9.5 ± 4.3  19.6 ± 9.1  AAT-1108 1079 7.6 ± 5.1  26.1 ± 2.7  AAT-1113 1084 91.1 ± 2.1   85.4 ± 7.8  AAT-1114 1085 56 ± 4.3  74.3 ± 4.8  AAT-1115 1086 13.1 ± 11.6  25.4 ± 21   AAT-1116 1087 5.8 ± 4.2  20.6 ± 13   AAT-1117 1088  15 ± 11.5   30 ± 18.1 AAT-1118 1089 6.6 ± 9.5  17.9 ± 10.8 AAT-1138 1109 36.3 ± 3.5    42 ± 5.4 AAT-1139 1110 27 ± 6.7  45.7 ± 3.4  AAT-1140 1111 100.4 ± 6.2    105.5 ± 3.6   AAT-1141 1112 16.7 ± 32.6    47 ± 34.4 AAT-1142 1113 18.9 ± 16.5  68.8 ± 12.2 AAT-1143 1114 10.1 ± 16    33.1 ± 21.4 AAT-1144 1115 19.7 ± 19.6  55.4 ± 31.2 AAT-1145 1116 31.4 ± 18.7  55.3 ± 26.9 AAT-1165 1136 41.6 ± 16.7  53.6 ± 15.3 AAT-1166 1137 12.5 ± 15.7  45.6 ± 19   AAT-1167 1138 76.7 ± 4.5    92 ± 5.4 AAT-1168 1139 29.2 ± 6     53.4 ± 17.4 AAT-1169 1140 57.7 ± 5     75.2 ± 5    AAT-1170 1141 2.8 ± 8.7  26.3 ± 21.9 AAT-1171 1142 2.2 ± 6.8  30.1 ± 10.2 AAT-1172 1143 9.5 ± 6.4  23.2 ± 16.1 AAT-1173 1144 21 ± 5.1  36.4 ± 2.8  AAT-1174 1145 10.5 ± 1.5   38.3 ± 10.5 AAT-1175 1146  9 ± 3.1 40.9 ± 1.4  AAT-1176 1147 16.4 ± 18.4    36 ± 25.5 AAT-1232 1203 18.8 ± 11.6  36.5 ± 9.2  AAT-1233 1204 2.2 ± 6.4  15.4 ± 5.5  AAT-1234 1205 3.8 ± 7    15.8 ± 7.1  AAT-1235 1206 3.7 ± 10.8 22.8 ± 20.1 AAT-1236 1207 6.3 ± 2.3  21.1 ± 11.2 AAT-1237 1208 7.6 ± 12.7 30.3 ± 31.2 AAT-1238 1209 52.8 ± 2.4   66.6 ± 9.6  AAT-1239 1210 9.1 ± 5.7  38.7 ± 8.6  AAT-1240 1211  15 ± 13.9 41.8 ± 23.3 AAT-1279 1250 8.7 ± 24.8 32.1 ± 24.3 AAT-1280 1251 11.3 ± 22.1  47.7 ± 19.8 AAT-1281 1252 56.5 ± 14.5  83.6 ± 15.6 AAT-1283 1254 69.8 ± 5.1   78.6 ± 7.8  AAT-1284 1255 89.3 ± 8     116.1 ± 5.9   AAT-1286 1257 6.6 ± 1.8  46.4 ± 5.7  AAT-1296 1119 85.8 ± 5.4   107.3 ± 5     77.8 ± 1.4 80.1 ± 3.4 AAT-1297 1120 85.6 ± 14.2  104.2 ± 11.6  94.4 ± 5.7 97.5 ± 3.5 AAT-1298 1121 94.6 ± 10.8  125.2 ± 7.3   80.7 ± 5.1   85 ± 6.5 AAT-1324 1295 49.8 ± 10    64.3 ± 13.8 AAT-1325 1296 38.5 ± 5.8     54 ± 10.6 AAT-1326 1297 6.7 ± 5.4  27.7 ± 9.4  AAT-1336 1307 19.5 ± 4.6   38.1 ± 15.1 AAT-1337 1308 34.1 ± 3.6   53.1 ± 3    AAT-1338 1309 78 ± 6.4  82.4 ± 2.6  AAT-1339 1310 8.4 ± 5.3  37.6 ± 14.8 AAT-1348 1319 22.6 ± 4.9    49 ± 5.3 AAT-1352 1323 19.9 ± 2.3   35.9 ± 15.2 AAT-1353 1324 3.4 ± 6.3  18.8 ± 8    AAT-1354 1180 37.9 ± 5.6   65.9 ± 12.6 64.8 ± 3.3 68.4 ± 5.5 AAT-1355 1181 30.3 ± 6.8   51.4 ± 9.7   52.5 ± 10.6 65.7 ± 4   AAT-1356 1182 39.7 ± 2.6   55.8 ± 5.6  53.4 ± 2.1 58.4 ± 5.7 AAT-1357 1183 22.5 ± 5.4   36.6 ± 3.1  34.3 ± 8   41.9 ± 5.5 AAT-1358 1184 22.7 ± 8.1   41.2 ± 10.2 33.6 ± 2.4 41.9 ± 4.3 AAT-1359 1185 46.2 ± 4.9   59.5 ± 8.1   24.7 ± 16.2  27.5 ± 15.8 AAT-1360 1186 7.7 ± 2.9  28.3 ± 8.3  19.9 ± 7.7 22.7 ± 4.6 AAT-1361 1187 11.1 ± 6      29 ± 1.2 14.5 ± 6   17.8 ± 9.9 AAT-1390 1361 55.2 ± 4.2   72.7 ± 9    AAT-1391 1362 18.9 ± 5.7    44 ± 8.8 AAT-1392 1363 15.8 ± 2.4   31.8 ± 8.1  AAT-1393 1364  66 ± 3.8  74.4 ± 2.7  AAT-1394 1365 54.5 ± 3.9   70.2 ± 7.5  AAT-1395 1366 11.7 ± 5.9   50.8 ± 8.4  AAT-1404 1375 45.9 ± 11.7  71.1 ± 12.5 AAT-1405 1376 47.3 ± 24.1  75.8 ± 30.3 AAT-1406 1377 11.6 ± 37    33.5 ± 21.1 AAT-1407 1378 24.1 ± 27.7  52.6 ± 26.2 AAT-1408 1379 10.4 ± 29.8  32.9 ± 25.7 AAT-1409 1380 56.7 ± 17.7  72.6 ± 20.1 AAT-1410 1381 75.9 ± 8.6   107.5 ± 5.6   AAT-1411 1382 18.4 ± 6.2   58.8 ± 11.5 AAT-1412 1383 27.7 ± 23.1  44.4 ± 16.4 AAT-1413 1384 48.7 ± 3.8   66 ± 6.3 AAT-1414 1385 62.3 ± 9.1   72.2 ± 7.8  AAT-1415 1386 62.6 ± 5.1   82.8 ± 3.5  AAT-1416 1387 2.6 ± 4.7  19.3 ± 2.7  AAT-1442 1413 5.6 ± 8.9  24.5 ± 8.6  AAT-1443 1414 3.4 ± 13.6 25.6 ± 16.4 AAT-1444 1415 12.3 ± 7.4   40.7 ± 11.7 AAT-1445 1416 2.5 ± 6    20.5 ± 11.8 AAT-1446 1417 12.8 ± 9.3   19.6 ± 16.4 AAT-1447 1418 8.5 ± 12   25.5 ± 13.5 AAT-1448 1419 2.4 ± 5.9  18.6 ± 14.5 AAT-1449 1420 2.2 ± 4.6  15.9 ± 6.8  AAT-1450 1421 2.1 ± 8.5  17.2 ± 6    AAT-1451 1422 4.5 ± 5.8  23.3 ± 7    AAT-1452 1423 26.9 ± 2.1   56.7 ± 12.5 AAT-1453 1424 3.5 ± 26   53.5 ± 29.2 AAT-1454 1425 4.7 ± 21.9 39.8 ± 22.4 AAT-1455 1426 7.7 ± 38.9 39.7 ± 43.7 AAT-1456 1427 7.4 ± 22.9 41.7 ± 20.3 AAT-1457 1428 7.5 ± 27.6 47.5 ± 29.3 AAT-1458 1429 6.1 ± 20.3 53.2 ± 7.5  AAT-1459 1430 3.2 ± 10.6 41.1 ± 19.7 AAT-1460 1431 6.3 ± 14.8 40.8 ± 5.9  AAT-1461 1432 2.5 ± 12.5 32.9 ± 8.8  AAT-1462 1433 1.8 ± 4.4  23.4 ± 14.5 AAT-1463 1434 1.9 ± 4.7  23.8 ± 10.3 AAT-1464 1435 1.9 ± 6.8  19.5 ± 23   AAT-1465 1436 1.7 ± 13.1 18.5 ± 14.5 AAT-1466 1437 9.3 ± 3.9  30.5 ± 3.5  AAT-1467 1438 56.6 ± 2.2   70.4 ± 3.6  AAT-1468 1439 6.2 ± 3.7  44.2 ± 4.3  AAT-1469 1440 3.7 ± 5.8  37.7 ± 8.2  AAT-1470 1441 3.2 ± 6.5  23.7 ± 4.2  AAT-1471 1442 1.6 ± 13.5   18 ± 31.3 AAT-1472 1443 1.3 ± 12.3 17.8 ± 15.1 AAT-1473 1444 2.8 ± 10.1 23.4 ± 10.2 AAT-1474 1445 4.5 ± 4.2  23.7 ± 4.2  AAT-1475 1446 11.1 ± 7.4   45.6 ± 10.6 AAT-1476 1447 7.1 ± 4.8  38.8 ± 5.9  AAT-1477 1448 2.8 ± 2.8  25.1 ± 9.6  AAT-1478 1449 5.2 ± 25.2 38.5 ± 24.7 AAT-1479 1450 13.3 ± 23.3  27.8 ± 11.5 AAT-1480 1451 35.7 ± 22.6  62.7 ± 29.2 AAT-1481 1452 5.4 ± 18.5   32 ± 18.6 AAT-1482 1453 6.9 ± 22.6 24.8 ± 20.7 AAT-1483 1454 12.4 ± 10.4    44 ± 21.5 AAT-1484 1455 50.9 ± 4.9     59 ± 10.9 AAT-1485 1456 73.7 ± 7.7   84.2 ± 2    AAT-1486 1457 13.7 ± 3.7   25.1 ± 8.4  AAT-1487 1458 11.7 ± 9.5   20.9 ± 9.1  AAT-1488 1459 3.5 ± 8.4  13.8 ± 5.8  AAT-1489 1460 3.3 ± 5    15.1 ± 7.1  AAT-1490 1461 10.2 ± 3.6   25.5 ± 13.5 AAT-1491 1462  10 ± 5.7  23.2 ± 2.6  AAT-1492 1463 23.8 ± 4.8   42.2 ± 4    AAT-1493 1464 26.7 ± 4.5   44.3 ± 16.9 AAT-1494 1465 30.4 ± 3.7   37.1 ± 4.5  AAT-1495 1466 7.1 ± 4.2  19.4 ± 13.2 AAT-1496 1467 17.6 ± 4.7   29.6 ± 2.5  AAT-1497 1468 17.3 ± 5.1   21.6 ± 10.9 AAT-1499 1470 106.7 ± 10.4   81.1 ± 5    AAT-1501 1472 38 ± 2.1  38.7 ± 2.9  AAT-1502 1473 10.5 ± 5     26.5 ± 6.1  AAT-1503 1474 39.4 ± 7.1   52.6 ± 14.8 AAT-1504 1475 9.1 ± 6.3  26.6 ± 17.1 AAT-1505 1476 7.4 ± 12   20.3 ± 9.8  AAT-1506 1477 60.9 ± 5.5   81.5 ± 2.5  AAT-1507 1478 3.9 ± 10.2 23.4 ± 10.9 AAT-1508 1479 5.4 ± 16.3 21.5 ± 22   AAT-1509 1480 12.7 ± 10.6  29.2 ± 25   AAT-1510 1481 5.7 ± 2.4  25.4 ± 27   AAT-1511 1482 15.7 ± 4.6    24 ± 8.5 AAT-1512 1483 57.5 ± 4.2   58.6 ± 7.8  AAT-1513 1484  6 ± 3.2 19.1 ± 7.2  AAT-1514 1485 21.7 ± 7.2   28.8 ± 10.7 AAT-1515 1486 72.4 ± 1.1   71.8 ± 0.6  AAT-1516 1487 18.3 ± 4.5   29.9 ± 8    AAT-1517 1488 6.6 ± 1    24.8 ± 2.4  AAT-2872 122.6 ± 2.1    21.8 ± 15.2 AAT-2880 104.5 ± 7.5    20.2 ± 16.1 AAT-3167 96.3 ± 2.3   20.8 ± 8.4  AAT-3169 90.4 ± 4.9   18.4 ± 5.9  AAT-3170 86.8 ± 8.8   25.7 ± 12.5 AAT-3172 93 ± 1.8  20.2 ± 3.2  AAT-3175 85.6 ± 1.6   15.1 ± 3.5  AAT-3180 90.2 ± 4.3   23.8 ± 5    AAT-3181 114.4 ± 4.1    28.7 ± 6.1  AAT-3182 114.2 ± 0.8    31.1 ± 2.6  109.1 ± 7.2  102.4 ± 5.6 

Example 3: DsiRNA Inhibition of α-1 Antitrypsin—Secondary Screen

96 asymmetric DsiRNAs (96 targeting Hs α-1 antitrypsin, 7 of which also targeted Mm α-1 antitrypsin) of the above experiment were then examined in a secondary assay (“Phase 2”), with results of such assays presented in histogram form in FIGS. 3A to 3H. Specifically, the 96 asymmetric DsiRNAs selected from those tested above were assessed for inhibition of human α-1 antitrypsin at 1 nM, 0.1 nM and 0.03 nM in the environment of human Huh7 cells (FIGS. 3A to 3D). These 96 asymmetric DsiRNAs were also assessed for inhibition of mouse α-1 antitrypsin at 1 nM, 0.1 nM and 0.03 nM in the environment of mouse AML12 cells (FIGS. 3E to 3H). As shown in FIGS. 3A to 3D, most asymmetric DsiRNAs reproducibly exhibited significant human α-1 antitrypsin inhibitory efficacies at sub-nanomolar concentrations when assayed in the environment of Huh7 cells. In addition, as shown in FIGS. 3E to 3H, a limited number of asymmetric DsiRNAs were identified to possess significant mouse α-1 antitrypsin inhibitory efficacies at sub-nanomolar concentrations when assayed in the environment of mouse AML12 cells.

Example 4: Modified Forms of α-1 Antitrypsin-Targeting DsiRNAs Reduced α-1 Antitrypsin Levels In Vitro

Twenty-four α-1 antitrypsin-targeting DsiRNAs of the above initial screen (AAT-486, -490, -496, -508, -810, -898, -899, -911, -1069, -1360, -1361, -1449, -1450, -1453, -1458, -1459, -1460, -1461, -1462, -1463, -1465, -1471, -1476 and -1477) were prepared with 2′-O-methyl guide and passenger strand modification patterns as shown in the schematics of FIG. 4A (with individual strand modification patterns shown in isolation in FIG. 4B; modifications included “M107” modified passenger strands and above-described guide strand modification patterns “M8”, “M17”, “M35” and “M48”). For each of the twenty-four DsiRNA sequences, DsiRNAs possessing each of the four guide strand modification patterns M8, M17, M35 and M48 were assayed for α-1 antitrypsin inhibition in human Huh7 cells at 1.0 nM, 0.1 nM and 0.03 nM concentrations in the environment of the Huh7 cells. Results of these experiments are presented as histograms in FIGS. 4C to 4F. In general, the twenty-four DsiRNA sequences exhibited a trend towards reduced efficacy of α-1 antitrypsin inhibition as the extent of 2′-O-methyl modification of the guide strand increased. However, for all DsiRNA sequences examined, a modification pattern could be identified that allowed the DsiRNA to retain significant α-1 antitrypsin inhibitory efficacy in vitro. It was also notable that a number of these DsiRNAs (e.g., AAT-486, AAT-490, AAT-899, AAT-911, AAT-1361, AAT-1458, AAT-1459, AAT-1460, AAT-1462, AAT-1463, AAT-1465 and AAT-1477) exhibited robust α-1 antitrypsin inhibitory efficacy in even the most highly modified states examined. Thus, such highly active modified DsiRNA sequences possessed modification patterns believed to be capable of stabilizing such DsiRNAs and/or reducing immunogenicity of such DsiRNAs when therapeutically administered to a subject in vivo.

Example 5: Additional Forms of α-1 Antitrypsin-Targeting DsiRNAs Possessing Modifications of Both Guide and Passenger Strands Reduced α-1 Antitrypsin Levels In Vitro

Sixteen α-1 antitrypsin-Targeting DsiRNAs of the above experiments (AAT-490, AAT-496, AAT-810, AAT-898, AAT-899, AAT-1360, AAT-1361, AAT-1450, AAT-1453, AAT-1458, AAT-1459, AAT-1461, AAT-1462, AAT-1463, AAT-1471 and AAT-1477) were prepared with 2′-O-methyl passenger strand and guide strand modification patterns as represented above and in FIGS. 5A to 5C (including passenger strand modification patterns “SM14”, “SM24”, “SM107”, “SM250”, “SM251” and “SM252” and guide strand modification patterns “M48”, “M8” and “M17”). For each of the sixteen DsiRNA sequences, DsiRNAs possessing each of the six passenger strand modification patterns M14, M24, M107, M250, M251 and M252 and one preferred guide strand modification pattern (selected from among guide strand modification patterns M48, M8 and M17) were assayed for α-1 antitrypsin inhibition in human Huh7 cells at 1.0 nM, 0.1 nM and 0.03 nM (30 picomolar) concentrations in the environment of the Huh7 cells. Results of these experiments are presented as histograms in FIGS. 5D to 5H. For all DsiRNA sequences examined, at least one duplex possessing extensive modification of both guide and passenger strands could be identified that allowed the DsiRNA to retain significant α-1 antitrypsin inhibitory efficacy in vitro. It was notable that many of these DsiRNAs (e.g., AAT-898, AAT-899, AAT-1461, AAT-1462, AAT-1463, AAT-1471 and AAT-1477) exhibited robust α-1 antitrypsin inhibitory efficacy across all modified states examined. Such highly active modified DsiRNA sequences possess modification patterns believed to be capable of stabilizing such DsiRNAs and/or reducing immunogenicity of such DsiRNAs when therapeutically administered to a subject in vivo.

AAT-490-M250-ExM8 (SEQ ID NO: 2077) 5′-UCCAACAGCACCAAUAUCUUCUUct-3′ (SEQ ID NO: 3493) 3′-UCAGGUUGUCGUGGUUAUAGAAGAAGAU

GCU

UCGT-5′ AAT-899-M252-ExM17 (SEQ ID NO: 73) 5′-UUUUUGCUCUGGUGAAUUACAUCtt-3′ (SEQ ID NO: 3494) 3′-UCAAAAACGAGACCACUUAAUGUAGAAU

GCU

UCGT-5′ AAT-1361-M250-ExM48 (SEQ ID NO: 134) 5′-AGGCUGUGCUGACCAUCGACGAGaa-3′ (SEQ ID NO: 3495) 3′-AUUCCGACACGACUGGUAGCUGCUCUUU

GCU

UCGT-5′ AA T-1462-M24-ExM17 (SEQ ID NO: 1787) 5′-CCCUUUGUCUUCUUAAUGAUUGAac-3′ (SEQ ID NO: 3496) 3′-UUGGGAAACAGAAGAAUUACUAACUUGU

GCU

UCGT-5′ AAT-1462-M252-ExM17 (SEQ ID NO: 1787) 5′-CCCUUUGUCUUCUUAAUGAUUGAac-3′ (SEQ ID NO: 3496) 3′-UUGGGAAACAGAAGAAUUACUAACUUGU

GCU

UCGT-5′ AAT-1463-M251-ExM48 (SEQ ID NO: 1788) 5′-CCUUUGUCUUCUUAAUGAUUGAAca-3′ (SEQ ID NO: 3497) 3′-UGGGAAACAGAAGAAUUACUAACUUGUU

GCU

UCGT-5′ AAT-1471-M250-ExM48 (SEQ ID NO: 1796) 5′-UUCUUAAUGAUUGAACAAAAUACca-3′ (SEQ ID NO: 3498) 3′-AGAAGAAUUACUAACUUGUUUUAUGGUU

GCU

UCGT-5′ AAT-1477-M251-ExM48 (SEQ ID NO: 1802) 5′-AUGAUUGAACAAAAUACCAAGUCtc-3′ (SEQ ID NO: 3499) 3′-AUUACUAACUUGUUUUAUGGUUCAGAGU

GCU

UCGT-5′ where an underscore indicates a 2′-O-methyl RNA and “A” in bold, italics indicates a 2′-Fluoro-adenine residue. The above “single-strand-extended” (or “ss-extended”) forms of the dsRNAs of the invention were also tested for α-1 antitrypsin knockdown activity in Huh7 cells, and as shown in FIG. 6B, all exhibited robust α-1 antitrypsin inhibitory efficacy, with all measured IC₅₀ values in the 0.1 to 20 pM range, and the lowest measured IC₅₀ value of 0.1 pM obtained for the AAT-1477-M251-ExM48 extended dsNA (referred to as “SERPINA1 1477-M251-ExM48” in FIG. 6B). Each of these extended forms of dsNA is also expected to exhibit a lack of immunogenicity (e.g., effectively no induction of IFN and/or PKR effect(s)) when administered in vivo and/or in an in vitro model or assay system for predicting or assessing in vivo immunogenicity, likely attributable to the masking provided by the modifications present within the single-strand extensions of these dsRNA sequences. Such dsRNA sequences possess modification patterns believed to be capable of stabilizing such dsRNAs and/or reducing immunogenicity of such dsRNAs—even for dsRNAs of such length—when therapeutically administered to a subject in vivo.

Example 7: Assessment of In Vivo Efficacy of α-1 Antitrypsin-Targeting DsiRNAs

The ability of certain, active α-1 antitrypsin-targeting DsiRNAs to reduce α-1 antitrypsin levels within a mouse, optionally a mouse model of liver disease is examined. Animals are randomized and assigned to groups based on marker levels. Dosing of animals with lipid nanoparticles (LNPs) containing DsiRNAs (optionally, an LNP formulation named EnCore-2072 is employed) is initiated on day 0. Animals are dosed at 5 mg/kg iv, tiw×2 (6 doses total). Animals are sacrificed 48 hrs after the last dose. Liver is dissected and weighed, and α-1 antitrypsin levels are assessed (optionally, for a mouse model of a liver disease or disorder, the extent of reduction and/or prevention of the disease or disorder is assessed).

Example 8: Indications

The present body of knowledge in α-1 antitrypsin research indicates the need for methods to assay α-1 antitrypsin activity and for compounds that can regulate α-1 antitrypsin expression for research, diagnostic, and therapeutic use. As described herein, the nucleic acid molecules of the present invention can be used in assays to diagnose disease state related to α-1 antitrypsin levels. In addition, the nucleic acid molecules can be used to treat disease state related to α-1 antitrypsin misregulation, levels, etc.

Particular disorders and disease states that can be associated with α-1 antitrypsin expression modulation include, but are not limited to chronic liver disease, liver inflammation, cirrhosis, liver fibrosis and hepatocellular carcinoma.

Other therapeutic agents can be combined with or used in conjunction with the nucleic acid molecules (e.g. DsiRNA molecules) of the instant invention. Those skilled in the art will recognize that other compounds and therapies used to treat the diseases and conditions described herein can be combined with the nucleic acid molecules of the instant invention (e.g. siNA molecules) and are hence within the scope of the instant invention. For example, for combination therapy, the nucleic acids of the invention can be prepared in one of at least two ways. First, the agents are physically combined in a preparation of nucleic acid and other agent, such as a mixture of a nucleic acid of the invention encapsulated in liposomes and other agent in a solution for intravenous administration, wherein both agents are present in a therapeutically effective concentration (e.g., the other agent in solution to deliver 1000-1250 mg/m2/day and liposome-associated nucleic acid of the invention in the same solution to deliver 0.1-100 mg/kg/day). Alternatively, the agents are administered separately but simultaneously or successively in their respective effective doses (e.g., 1000-1250 mg/m2/d other agent and 0.1 to 100 mg/kg/day nucleic acid of the invention).

Example 9: Serum Stability for DsiRNAs

Serum stability of DsiRNA agents is assessed via incubation of DsiRNA agents in 50% fetal bovine serum for various periods of time (up to 24 h) at 37° C. Serum is extracted and the nucleic acids are separated on a 20% non-denaturing PAGE and can be visualized with Gelstar stain. Relative levels of protection from nuclease degradation are assessed for DsiRNAs (optionally with and without modifications).

Example 10: Diagnostic Uses

The DsiRNA molecules of the invention can be used in a variety of diagnostic applications, such as in the identification of molecular targets (e.g., RNA) in a variety of applications, for example, in clinical, industrial, environmental, agricultural and/or research settings. Such diagnostic use of DsiRNA molecules involves utilizing reconstituted RNAi systems, for example, using cellular lysates or partially purified cellular lysates. DsiRNA molecules of this invention can be used as diagnostic tools to examine genetic drift and mutations within diseased cells. The close relationship between DsiRNA activity and the structure of the target α-1 antitrypsin RNA allows the detection of mutations in a region of the α-1 antitrypsin molecule, which alters the base-pairing and three-dimensional structure of the target α-1 antitrypsin RNA. By using multiple DsiRNA molecules described in this invention, one can map nucleotide changes, which are important to RNA structure and function in vitro, as well as in cells and tissues. Cleavage of target α-1 antitrypsin RNAs with DsiRNA molecules can be used to inhibit gene expression and define the role of specified gene products in the progression of a α-1 antitrypsin-associated disease or disorder. In this manner, other genetic targets can be defined as important mediators of the disease. These experiments will lead to better treatment of the disease progression by affording the possibility of combination therapies (e.g., multiple DsiRNA molecules targeted to different genes, DsiRNA molecules coupled with known small molecule inhibitors, or intermittent treatment with combinations of DsiRNA molecules and/or other chemical or biological molecules). Other in vitro uses of DsiRNA molecules of this invention are well known in the art, and include detection of the presence of RNAs associated with a disease or related condition. Such RNA is detected by determining the presence of a cleavage product after treatment with a DsiRNA using standard methodologies, for example, fluorescence resonance emission transfer (FRET).

In a specific example, DsiRNA molecules that cleave only wild-type or mutant or polymorphic forms of the target α-1 antitrypsin RNA are used for the assay. The first DsiRNA molecules (i.e., those that cleave only wild-type forms of target α-1 antitrypsin RNA) are used to identify wild-type α-1 antitrypsin RNA present in the sample and the second DsiRNA molecules (i.e., those that cleave only mutant or polymorphic forms of target RNA) are used to identify mutant or polymorphic α-1 antitrypsin RNA in the sample. As reaction controls, synthetic substrates of both wild-type and mutant or polymorphic α-1 antitrypsin RNA are cleaved by both DsiRNA molecules to demonstrate the relative DsiRNA efficiencies in the reactions and the absence of cleavage of the “non-targeted” α-1 antitrypsin RNA species. The cleavage products from the synthetic substrates also serve to generate size markers for the analysis of wild-type and mutant α-1 antitrypsin RNAs in the sample population. Thus, each analysis requires two DsiRNA molecules, two substrates and one unknown sample, which is combined into six reactions. The presence of cleavage products is determined using an RNase protection assay so that full-length and cleavage fragments of each α-1 antitrypsin RNA can be analyzed in one lane of a polyacrylamide gel. It is not absolutely required to quantify the results to gain insight into the expression of mutant or polymorphic α-1 antitrypsin RNAs and putative risk of α-1 antitrypsin-associated phenotypic changes in target cells. The expression of α-1 antitrypsin mRNA whose protein product is implicated in the development of the phenotype (i.e., disease related/associated) is adequate to establish risk. If probes of comparable specific activity are used for both transcripts, then a qualitative comparison of α-1 antitrypsin RNA levels is adequate and decreases the cost of the initial diagnosis. Higher mutant or polymorphic form to wild-type ratios are correlated with higher risk whether α-1 antitrypsin RNA levels are compared qualitatively or quantitatively.

All patents and publications mentioned in the specification are indicative of the levels of skill of those skilled in the art to which the invention pertains. All references cited in this disclosure are incorporated by reference to the same extent as if each reference had been incorporated by reference in its entirety individually.

One skilled in the art would readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. The methods and compositions described herein as presently representative of preferred embodiments are exemplary and are not intended as limitations on the scope of the invention. Changes therein and other uses will occur to those skilled in the art, which are encompassed within the spirit of the invention, are defined by the scope of the claims.

It will be readily apparent to one skilled in the art that varying substitutions and modifications can be made to the invention disclosed herein without departing from the scope and spirit of the invention. Thus, such additional embodiments are within the scope of the present invention and the following claims. The present invention teaches one skilled in the art to test various combinations and/or substitutions of chemical modifications described herein toward generating nucleic acid constructs with improved activity for mediating RNAi activity. Such improved activity can comprise improved stability, improved bioavailability, and/or improved activation of cellular responses mediating RNAi. Therefore, the specific embodiments described herein are not limiting and one skilled in the art can readily appreciate that specific combinations of the modifications described herein can be tested without undue experimentation toward identifying DsiRNA molecules with improved RNAi activity.

The invention illustratively described herein suitably can be practiced in the absence of any element or elements, limitation or limitations that are not specifically disclosed herein. Thus, for example, in each instance herein any of the terms “comprising”, “consisting essentially of”, and “consisting of” may be replaced with either of the other two terms. The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments, optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the description and the appended claims.

In addition, where features or aspects of the invention are described in terms of Markush groups or other grouping of alternatives, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group or other group.

The use of the terms “a” and “an” and “the” and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to,”) unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

Embodiments of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of those embodiments may become apparent to those of ordinary skill in the art upon reading the foregoing description.

The inventors expect skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims. 

We claim:
 1. A method for reducing expression of a target α-1 antitrypsin mRNA in a mammal, the method comprising administering a double stranded nucleic acid (dsNA) to said mammal in an amount sufficient to reduce expression of said target α-1 antitrypsin mRNA in said mammal, wherein said dsNA comprises a first nucleic acid strand and a second nucleic acid strand that each have a nucleotide sequence consisting of 21 nucleotides, wherein said first nucleic acid strand consists of the nucleic acid sequence of SEQ ID NO:1908, wherein said second nucleic acid strand is 100% complementary to SEQ ID NO: 1908, and wherein one or more of said 21 nucleotides of at least one of said first and second nucleic acid strands are modified nucleotides.
 2. The method of claim 1, wherein said administering step comprises a mode selected from the group consisting of intravenous injection, intramuscular injection, intraperitoneal injection, infusion, subcutaneous injection, transdermal, aerosol, rectal, vaginal, topical, oral and inhaled delivery.
 3. The method of claim 1, wherein said mammal is human.
 4. The method of claim 1, wherein said one or more modified nucleotides is selected from: a 2′-O-methyl nucleotide; a 2′-fluoro nucleotide; and a 2′-O-methyl nucleotide and a 2′-fluoro nucleotide.
 5. The method of claim 4, wherein said dsNA comprises one or more phosphorothioate linkages.
 6. The method of claim 5, wherein said dsNA comprises a 3′-terminus having one or more phosphorothioate linkages.
 7. The method of claim 1, wherein said first or said second nucleic acid strand comprises one or more phosphorothioate linkages or each of said first and second nucleic acid strands comprises one or more phosphorothioate linkages.
 8. The method of claim 7, wherein each of said first and second nucleic acid strands comprises a 3′ terminus, and wherein said first nucleic acid strand comprises a phosphorothioate linkage at said 3′ terminus or said second nucleic acid strand comprises a phosphorothioate linkage at said 3′-terminus or each of said first and second nucleic acid strands comprises a phosphorothioate linkage at said 3′-terminus.
 9. The method of claim 1, wherein said dsNA comprises a targeting ligand attached to said first nucleic acid strand, said second nucleic acid strand, or both.
 10. The method of claim 9, wherein said targeting ligand comprises a GalNAc moiety or a tri-antennary GalNAc moiety.
 11. The method of claim 1, wherein said first or second nucleic acid strand comprises one or more inverted abasic residues or each of said first and second nucleic acid strands comprises one or more inverted abasic residues.
 12. The method of claim 11, wherein said one or more modified nucleotides is selected from: a 2′-O-methyl nucleotide, a 2′-fluoro nucleotide; and a 2′-O-methyl nucleotide and a 2′-fluoro nucleotide.
 13. The method of claim 12, wherein said dsNA comprises one or more phosphorothioate linkages.
 14. The method of claim 13, wherein each of said first and second nucleic acid strands comprises a 3′ terminus, and wherein said first nucleic acid strand comprises a phosphorothioate linkage at said 3′ terminus or said second nucleic acid strand comprises a phosphorothioate linkage at said 3′-terminus or each of said first and second nucleic acid strands comprises a phosphorothioate linkage at said 3′-terminus.
 15. The method of claim 14, wherein said dsNA comprises a targeting ligand attached to said first nucleic acid strand, said second nucleic acid strand, or both.
 16. The method of claim 15, wherein said targeting ligand comprises a GalNAc moiety or a tri-antennary GalNAc moiety.
 17. The method of claim 9, wherein said first or second nucleic acid strand comprises one or more inverted abasic residues or each of said first and second nucleic acid strands comprises one or more inverted abasic residues.
 18. The method of claim 1, wherein a 3′-terminus of said first nucleic acid strand and a 5′-terminus of said second strand form a blunt end.
 19. The method of claim 1, wherein termini of said first and second nucleic acid strands each form a blunt end.
 20. The method of claim 1, wherein said first and second nucleic acid strands are separate nucleic acid molecules.
 21. The method of claim 1, wherein each of said 21 nucleotides of each of said first and second nucleic acid strands is a modified nucleotide.
 22. The method of claim 1, wherein said dsNA comprises one or more modifications patterns.
 23. The method of claim 9, wherein said first and second nucleic acid strands are separate nucleic acid molecules.
 24. The method of claim 9, wherein each of said 21 nucleotides of each of said first and second nucleic acid strands is a modified nucleotide.
 25. The method of claim 9, wherein said dsNA comprises one or more modifications patterns. 